Pyridine-substituted benzanilides as potassium ion channel openers

ABSTRACT

The present invention provides a genus of pyridine-substituted benzanilides that are useful as openers of potassium ion channels. The compounds of the invention are of use in both therapeutic and diagnostic methods.

CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] The present application claims priority to U.S. Provisionalpatent application Ser. No. 60/147,221, filed on Aug. 4, 1999, and is acontinuation-in-part of U.S. patent application Ser. No. 09/632,576,filed on Aug. 4, 2000 the disclosure of each of which is incorporatedherein by reference in its entirety for all purposes.

FIELD OF THE INVENTION

[0002] This invention relates to the use of certain pyridine-substitutedbenzanilide derivatives as potassium channel openers and to thetreatment of diseases modulated by potassium channels. Additionally,this invention relates to novel compounds that are useful as potassiumchannel openers.

BACKGROUND OF THE INVENTION

[0003] Ion channels are cellular proteins that regulate the flow ofions, including calcium, potassium, sodium and chloride, into and out ofcells. These channels are present in all human cells and affect suchprocesses as nerve transmission, muscle contraction and cellularsecretion. Among the ion channels, potassium channels are the mostubiquitous and diverse, being found in a variety of animal cells such asnervous, muscular, glandular, immune, reproductive, and epithelialtissue. These channels allow the flow of potassium in and/or out of thecell under certain conditions. For example, the outward flow ofpotassium ions upon opening of these channels makes the interior of thecell more negative, counteracting depolarizing voltages applied to thecell. These channels are regulated, e.g., by calcium sensitivity,voltage-gating, second messengers, extracellular ligands, andATP-sensitivity.

[0004] Potassium channels have now been associated with a number ofphysiological processes, including regulation of heartbeat, dilation ofarteries, release of insulin, excitability of nerve cells, andregulation of renal electrolyte transport. Potassium channels are madeby alpha subunits that fall into at least 8 families, based on predictedstructural and functional similarities (Wei et al., Neuropharmacology35(7): 805-829 (1997)). Three of these families (Kv, eag-related, andKQT) share a common motif of six transmembrane domains and are primarilygated by voltage. Two other families, CNG and SK/IK, also contain thismotif but are gated by cyclic nucleotides and calcium, respectively. Thethree other families of potassium channel alpha subunits have distinctpatterns of transmembrane domains. Slo family potassium channels, or BKchannels have seven transmembrane domains (Meera et al., Proc. NatLAcad. Sci. U.S.A. 94(25): 14066-71 (1997)) and are gated by both voltageand calcium or pH (Schreiber et al., J Biol. Chem. 273: 3509-16 (1998)).Another family, the inward rectifier potassium channels (Kir), belong toa structural family containing two transmembrane domains, and an eighthfunctionally diverse family (TP, or “two-pore”) contains two tandemrepeats of this inward rectifier motif.

[0005] Potassium channels are typically formed by four alpha subunits,and can be homomeric (made of identical alpha subunits) or heteromeric(made of two or more distinct types of alpha subunits). In addition,potassium channels made from Kv, KQT and Slo or BK subunits have oftenbeen found to contain additional, structurally distinct auxiliary, orbeta, subunits. These subunits do not form potassium channelsthemselves, but instead they act as auxiliary subunits to modify thefunctional properties of channels formed by alpha subunits. For example,the Kv beta subunits are cytoplasmic and are known to increase thesurface expression of Kv channels and/or modify inactivation kinetics ofthe channel (Heinemann et al., J Physiol. 493: 625-633 (1996); Shi etal., Neuron 16(4): 843-852 (1996)). In another example, the KQT familybeta subunit, minK, primarily changes activation kinetics (Sanguinettiet al., Nature 384: 80-83 (1996)).

[0006] Slo or BK potassium channels are large conductance potassiumchannels found in a wide variety of tissues, both in the central nervoussystem and periphery. They play a key role in the regulation ofprocesses such as neuronal integration, muscular contraction and hormonesecretion. They may also be involved in processes such as lymphocytedifferentiation and cell proliferation, spermatocyte differentiation andsperm motility. Three alpha subunits of the Slo family have been cloned,i.e., Slo1, Slo2, and Slo3 (Butler et al., Science 261: 221-224 (1993);Schreiber et al., J Biol. Chem., 273: 3509-16 (1998); and Joiner et al.,Nature Neurosci. 1: 462-469 (1998)). These Slo family members have beenshown to be voltage and/or calcium gated, and/or regulated byintracellular pH.

[0007] Certain members of the Kv family of potassium channels wererecently renamed (see Biervert, et al., Science 279: 403-406 (1998)).KvLQTl was re-named KCNQ1, and the KvLQT1-related channels (KvLR1 andKvLR2) were renamed KCNQ2 and KCNQ3, respectively. More recently, afourth member of the KCNQ subfamily was identified (KCNQ4) as a channelexpressed in sensory outer hair cells (Kubisch, et al., Cell 96(3):437-446 (1999)).

[0008] KCNQ2 and KCNQ3 have been shown to be nervous system-specificpotassium channels associated with benign familial neonatal convulsions(“BFNC”), a class of idiopathic generalized epilepsy (see, Leppert, etal., Nature 337: 647-648 (1989)). These channels have been linked toM-current channels (see, Wang, et al, Science 282: 1890-1893 (1998)).The discovery and characterization of these channels and currentsprovides useful insights into how these voltage dependent (Kv) potassiumchannels function in different environments, and how they respond tovarious activation mechanisms. Such information has now led to theidentification of modulators of KCNQ2 and KCNQ3 potassium channels orthe M-current, and the use of such modulators as therapeutic agents. Themodulators are the subject of the present invention.

[0009] KCNQ4 (Kubsich et al., Cell 96(3): 437 (1999)), KCNQ5 (Kananuraet aL, Neuroreport 11 (9):2063 (2000)), KCNQ 3/5 (Wickenden et al., Br.J. Pharma 132: 381 (2001)) and KCNQ6 have also recently been described.

SUMMARY OF THE INVENTION

[0010] The present invention provides pyridine-substituted benzanilidecompounds, and phannaceutically acceptable salts thereof (“compounds ofthe invention”), which are useful in the treatment of diseases throughthe modulation of potassium ion flux through voltage-dependent potassiumchannels.

[0011] In one aspect, the present invention provides compounds having astructure according to Formula I:

[0012] in which the symbol Ar¹ represents a member selected from thegroup consisting of aryl, substituted aryl, heteroaryl and substitutedheteroaryl. The letter X represents a member selected from the groupconsisting of O, S and N-R¹, in which R¹ is H, (C₁-C₈)alkyl, substituted(C₁-C₈)alkyl, heteroalkyl, substituted heteroalkyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, aryl(C₁-C4)alkyl, substitutedaryl(C₁-C₄)alkyl, CN, —C(O)R², —OR³, —C(O)NR³R⁴, or —S(O)₂NR³R⁴. Thesymbol R² represents a member selected from the group consisting of(C₁-C₈)alkyl, substituted (Cl-C8)alkyl, cycloalkyl, substitutedcycloalkyl, heteroalkyl, substituted heteroalkyl, heterocyclyl,substituted heterocyclyl, substituted aryl, heteroaryl, substitutedheteroaryl, aryl(C₁-C₄)alkyl and substituted aryl(C₁-C₄)alkyl. R³ and R⁴are each members independently selected from the group consisting ofhydrogen, (C₁-C₈)alkyl, substituted (C₁-C₈)alkyl, cycloalkyl,substituted cycloalkyl, heteroalkyl, substituted heteroalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, aryl(C₁-C₄)alkyl and substitutedaryl(C₁-C₄)alkyl. Alternatively, R³ and R⁴ can be combined with thenitrogen to which each is attached to form a 5-, 6- or 7-membered ring,optionally having additional heteroatoms at the ring vertices. Theletter Y represents a member selected from the group consisting ofhalogen, C₁-C₄ alkyl, C₁-C₄ substituted alkyl, —OCH₃ and 13 OCF₃.

[0013] In another aspect, the present invention provides pharmaceuticalcompositions comprising a pharmaceutically acceptable excipient and acompound having a structure according to Formula II:

[0014] in which the symbols Ar¹ and Ar2 independently represent aryl,substituted aryl, heteroaryl and substituted heteroaryl. The letterrepresents 0, S or N—R¹, in which R¹ is a H, (C₁-C₈)alkyl, substituted(C₁-C₈)alkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, aryl(C₁-C₄)alkyl, substituted aryl(C₁-C₄)alkyl, CN, —C(O)R²,—OR³, —C(O)NR³R⁴, or —S(O)₂NR³R⁴. The symbol R² represents (C₁-C₈)alkyl,substituted (C₁-C₈)alkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, aryl(C₁-C₄)alkyl or substitutedaryl(C₁-C₄)alkyl. R³ and R⁴ are each members independently selected fromthe group consisting of hydrogen, (C₁-C₈)alkyl, substituted(C₁-C₈)alkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, aryl(C₁-C₄)alkyl and substituted aryl(C₁-C₄)alkyl.Alternatively, R³ and R⁴ can be combined with the nitrogen to which eachis attached to form a 5-, 6- or 7-membered ring optionally havingadditional heteroatoms at the ring vertices.

[0015] In a further aspect, the present invention provides compoundshaving a structure according to Formula III:

[0016] In Formula III, the symbols Y¹ and y² independently represent H,methyl, methoxy, trifluoromethoxy, —CF₃ or halo, with the proviso thatboth y¹ and y² are not H. The symbols V and X independently represent H,halo, substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, NO₂, CN, CF₃, C(O)NR¹¹R¹² and C(O)R¹³.The symbols R¹¹, R¹² and R¹³ independently represent substituted orunsubstituted lower alkyl, substituted or unsubstituted lowerheteroalkyl, substituted or unsubstituted carbocycle, substituted orunsubstituted heterocycle, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. R¹¹ and R¹² can be joined intoa ring. Q and W independently represent —(CR₂R₃)_(t)—(CH₂)_(n)—,-(CH₂)_(n)—(CR₂R₃)_(t), —C(R⁴)═C(R⁵)—, or —C≡C— wherein R² and R³ areindependently F, substituted or unsubstituted lower alkyl or substitutedor unsubstituted lower heteroalkyl, wherein R² and R³ are optionallyjoined to form a cyclic structure which is a member selected from thegroup consisting of cycloalkyls and heterocycles, or R² and R³ togetherwith the carbon to which they are attached form carbonyl (i.e., —C(O)—).Q can optionally be a bond between the phenyl ring and Z, and W canoptionally be a bond between Z and R¹. The symbol Z represents —O—,—S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—,—N(R⁴)C(O)O—, and —SO₂N(R⁴)—, wherein R⁴ and R⁵ are membersindependently selected from the group consisting of H, substituted orunsubstituted lower alkyl, substituted or unsubstituted lowerheteroalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. R¹ is optionally joined together with either Xor R⁴ to form a substituted or unsubstituted heterocycle. The symbol mrepresents an integer from 0 to 2, inclusive; n represents and integerfrom 0 and 3, inclusive; and t represents an integer from 0 to 2,inclusive.

[0017] In yet another aspect, the present invention provides a methodfor modulating ion flux through voltage dependent potassium channels,comprising contacting a cell containing the target ion channels with acompound according to Formulae I-IV.

[0018] In still another aspect, the present invention provides a methodfor the treatment of diseases through modulation of ion flux throughvoltage dependent potassium channels, the method comprising treating thehost with an effective amount of a potassium channel opening compound ofFormula I-IV.

BRIEF DESCRIPTION OF THE DRAWINGS

[0019]FIG. 1 displays structures of representative compounds of theinvention.

DETAILED DESCRIPTION OF THE INVENTION AND THE PREFERRED EMBODIMENTS

[0020] Abbreviations and Definitions

[0021] The abbreviations used herein have their conventional meaningwithin the chemical and biological arts. For example: CHO, Chinesehamster ovary; EBSS, Earl's Balanced Salt Solution; KCNQ, potassiumchannel Q; KCNQ2, potassium channel Q2, hSK, Ca2+activated smallconductance potassium channels; SDS, sodium dodecyl sulfate; Et₃N,triethylamine; MeOH, methanol; and DMSO, dimethylsulfoxide.

[0022] Where substituent groups are specified by their conventionalchemical formulae, written from left to right, they equally encompassthe chemically identical substituents which would result from writingthe structure from right to left, e.g., —CH₂O— is intended to alsorecite —OCH₂—.

[0023] The term “alkyl,” by itself or as part of another substituent,means, unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e. C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude, but are not limited to, groups such as methyl, ethyl, n-propyl,isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, forexample, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Anunsaturated alkyl group is one having one or more double bonds or triplebonds. Examples of unsaturated alkyl groups include, but are not limitedto, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “alkyl,” unlessotherwise noted, is also meant to include those derivatives of alkyldefined in more detail below, such as “heteroalkyl.” Alkyl groups whichare limited to hydrocarbon groups are termed “homoalkyl”.

[0024] The term “alkylene” by itself or as part of another substituentmeans a divalent radical derived from an alkane, as exemplified, but notlimited, by —CH₂CH₂CH₂CH₂—, and further includes those groups describedbelow as heteroalkylene. Typically, an alkyl (or alkylene) group willhave from 1 to 24 carbon atoms, with those groups having 10 or fewercarbon atoms being preferred in the present invention. A “lower alkyl”or “lower alkylene” is a shorter chain alkyl or alkylene group,generally having eight or fewer carbon atoms.

[0025] The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy)are used in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

[0026] The term “heteroalkyl,” by itself or in combination with anotherterm, means, unless otherwise stated, a stable straight or branchedchain, or cyclic hydrocarbon radical, or combinations thereof,consisting of the stated number of carbon atoms and at least oneheteroatom selected from the group consisting of O, N, Si and S, andwherein the nitrogen and sulfur atoms may optionally be oxidized and thenitrogen heteroatom may optionally be quatemized. The heteroatom(s) 0, Nand S and Si may be placed at any interior position of the heteroalkylgroup or at the position at which the alkyl group is attached to theremainder of the molecule. Examples include, but are not limited to,—CH₂-CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃,—CH₂—CH₂,—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—)—CH₃, —Si(CH₃)₃,—CH₂—CH═N—OCH₃, and —CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may beconsecutive, such as, for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.Similarly, the term “heteroalkylene” by itself or as part of anothersubstituent means a divalent radical derived from heteroalkyl, asexemplified, but not limited by, —CH₂—CH₂—S—CH₂—CH₂— and—CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylene groups, heteroatoms can alsooccupy either or both of the chain termini (e.g., alkyleneoxy,alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Stillfurther, for alkylene and heteroalkylene linking groups, no orientationof the linking group is implied by the direction in which the formula ofthe linking group is written. For example, the formula —C(O)₂R′—represents both —C(O)₂R′— and —R′C(O)₂—.

[0027] The terms “cycloalkyl” and “heterocycloalkyl”, by themselves orin combination with other terms, represent, unless otherwise stated,cyclic versions of “alkyl” and “heteroalkyl”, respectively.Additionally, for heterocycloalkyl, a heteroatom can occupy the positionat which the heterocycle is attached to the remainder of the molecule.Examples of cycloalkyl include, but are not limited to, cyclopentyl,cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like.Examples of heterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofaran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

[0028] The terms “halo” or “halogen,” by themselves or as part ofanother substituent, mean, unless otherwise stated, a fluorine,chlorine, bromine, or iodine atom. Additionally, terms such as“haloalkyl,” are meant to include monohaloalkyl and polyhaloalkyl. Forexample, the term “halo(C₁-C₄)alkyl” is mean to include, but not belimited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

[0029] The term “aryl” means, unless otherwise stated, apolyunsaturated, aromatic, hydrocarbon substituent which can be a singlering or multiple rings (preferably from 1 to 3 rings) which are fusedtogether or linked covalently. The term “heteroaryl” refers to arylgroups (or rings) that contain from one to four heteroatoms selectedfrom N, O, and S, wherein the nitrogen and sulfiur atoms are optionallyoxidized, and the nitrogen atom(s) are optionally quatemized. Aheteroaryl group can be attached to the remainder of the moleculethrough a heteroatom. Non-limiting examples of aryl and heteroarylgroups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl,2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,5-thiazolyl, 2-fuiryl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituentsfor each of the above noted aryl and heteroaryl ring systems areselected from the group of acceptable substituents described below.

[0030] For brevity, the term “aryl” when used in combination with otherterms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl andheteroaryl rings as defined above. Thus, the term arylalkyl is meant toinclude those radicals in which an aryl group is attached to an alkylgroup (e.g., benzyl, phenethyl, pyridylmethyl and the like) includingthose alkyl groups in which a carbon atom (e.g., a methylene group) hasbeen replaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like).

[0031] Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl” and“heteroaryl”) are meant to include both substituted and unsubstitutedforms of the indicated radical. Preferred substituents for each type ofradical are provided below.

[0032] Substituents for the alkyl and heteroalkyl radicals (includingthose groups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′,—halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″,—NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —CN and—NO₂ in a number ranging from zero to (2m′+1), where m is the totalnumber of carbon atoms in such radical. R′, R″, R′″ and R″″ eachpreferably independently refer to hydrogen, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted aryl, e.g., aryl substitutedwith 1-3 halogens, substituted or unsubstituted alkyl, alkoxy orthioalkoxy groups, or arylalkyl groups. When a compound of the inventionincludes more than one R group, for example, each of the R groups isindependently selected as are each R′, R″, R′″ and R″″ groups when morethan one of these groups is present. When R′ and R″ are attached to thesame nitrogen atom, they can be combined with the nitrogen atom to forma 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant to include,but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the abovediscussion of substituents, one of skill in the art will understand thatthe term “alkyl” is meant to include groups including carbon atoms boundto groups other than hydrogen groups, such as haloalkyl (e.g., —CF₃ and—CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and thelike).

[0033] Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are varied and areselected from, for example: halogen, —OR′, ═O, ═NR, ═N—OR′, —NR′R″,—SR′, —halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″,—OC(O)NR′R″, —NR′—C(O)R, —NR′—C(O)NR″R′″, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —CN and —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy, andfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total number ofopen valences on the aromatic ring system; and where R′, R″, R′″ and R″″are preferably independently selected from hydrogen, (C₁-C₈)alkyl andheteroalkyl, unsubstituted aryl and heteroaryl, (unsubstitutedaryl)-(C₁-C₄)alkyl, and (unsubstituted aryl)oxy-(C₁-C₄)alkyl. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″and R″″ groups when more than one of these groups is present.

[0034] Two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —T—C(O)—(CRR′)_(q)—U—, wherein T and U are independently —NR—,—O—, —CRR′— or a single bond, and q is an integer of from 0 to 3.Alternatively, two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —A—(CH₂)_(r)—B—, wherein A and B are independently —CRR′—, —O—,—NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR— or a single bond, and r is aninteger of from 1 to 4. One of the single bonds of the new ring soformed may optionally be replaced with a double bond. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CRR′)_(s)—X—(CR″R′″)_(d)—, where s and d are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR—.The substituents R, R′, R″ and R′″ are preferably independently selectedfrom hydrogen or substituted or unsubstituted (C₁-C₆)alkyl.

[0035] As used herein, the term heteroatom is meant to include oxygen(O), nitrogen (N), sulfur (S) and silicon (Si).

[0036] The term “pharmaceutically acceptable salts” is meant to includesalts of the active compounds which are prepared with relativelynontoxic acids or bases, depending on the particular substituents foundon the compounds described herein. When compounds of the presentinvention contain relatively acidic functionalities, base addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, f-imaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge et a., “Pharmaceutical Salts”, Journal ofPharmaceutical Science, 1977, 66, 1-19). Certain specific compounds ofthe present invention contain both basic and acidic functionalities thatallow the compounds to be converted into either base or acid additionsalts.

[0037] The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

[0038] In addition to salt forms, the present invention providescompounds, which are in a prodrug form. Prodrugs of the compoundsdescribed herein are those compounds that readily undergo chemicalchanges under physiological conditions to provide the compounds of thepresent invention. Additionally, prodrugs can be converted to thecompounds of the present invention by chemical or biochemical methods inan ex vivo environment. For example, prodrugs can be slowly converted tothe compounds of the present invention when placed in a transdermalpatch reservoir with a suitable enzyme or chemical reagent.

[0039] Certain compounds of the present invention can exist inunsolvated forms as well as solvated forms, including hydrated forms. Ingeneral, the solvated forms are equivalent to unsolvated forms and areencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

[0040] Certain compounds of the present invention possess asymmetriccarbon atoms (optical centers) or double bonds; the racemates,diastereomers, geometric isomers and individual isomers are encompassedwithin the scope of the present invention.

[0041] The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine—125 (¹²⁵I) or carbon—14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

[0042] Introduction

[0043] The present invention provides compounds which, inter alia, areusefuil in the treatment of diseases through the modulation of potassiumion flux through voltage-dependent potassium channels. Moreparticularly, the invention provides compounds, compositions and methodsthat are useful in the treatment of central or peripheral nervous systemdisorders (e.g., migraine, ataxia, Parkinson's disease, bipolardisorders, trigeminal neuralgia, spasticity, mood disorders, braintumors, psychotic disorders, myokymia, seizures, epilepsy, hearing andvision loss, Alzheimer's disease, age-related memory loss, learningdeficiencies, anxiety and motor neuron diseases), and as neuroprotectiveagents (e.g., to prevent stroke and the like). Compounds of theinvention have use as agents for treating convulsive states, for examplethat following grand mal, petit mal, psychomotor epilepsy or focalseizure. The compounds of the invention are also useful in treatingdisease states such as gastroesophogeal reflux disorder andgastrointestinal hypomotility disorders.

[0044] Moreover, compounds of the invention are useful in the treatmentof pain, for example, neuropathic pain, inflammatory pain, cancer pain,migraine pain, and musculoskeletal pain. The compounds are also usefulto treat conditions, which may themselves be the origin of pain, forexample, inflammatory conditions, including arthritic conditions (e.g.,rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis and goutyarthritis) and non-articular inflammatory conditions (e.g., herniated,ruptured and prolapsed disc syndrome, bursitis, tendonitis,tenosynovitis, fibromyalgia syndrome, and other conditions associatedwith ligamentous sprain and regional musculoskeletal strain).Particularly preferred compounds of the invention are less ulcerogenicthan other anti-inflammatory agents (e.g., ibuprofen, naproxen andaspirin). Furthermore, the compounds of the invention are useful intreating conditions and pain associated with abnormally raised skeletalmuscle tone.

[0045] The compounds of the invention are also of use in treatinganxiety (e.g. anxiety disorders). Anxiety disorders are defined in theDiagnostic and Statistical Manual of Mental Disorders (ThirdEdition-revised 1987, published by the American Psychiatric Association,Washington, D.C., see, pages 235 to 253), as psychiatric conditionshaving symptoms of anxiety and avoidance behavior as characteristicfeatures. Included amongst such disorders are generalized anxietydisorder, simple phobia and panic disorder.

[0046] Anxiety also occurs as a symptom associated with otherpsychiatric disorders, for example, obsessive compulsive disorder,post-traumatic stress disorder, schizophrenia, mood disorders and majordepressive disorders, and with organic clinical conditions including,but not limited to, Parkinson's disease, multiple sclerosis, and otherphysically incapacitating disorders.

[0047] The development of therapeutic agents, which act on potassium ionchannels has received considerable recent attention. One group hasdescribed a family of N-alkyl benzamides that act by blocking potassiumchannels (see, PCT/US98/02364, published as WO 98/37068). In contrast,the benzanilides provided herein act by opening potassium channels.

[0048] In view of the above-noted discovery, the present inventionprovides compounds, compositions, and methods for increasing ion flux involtage-dependent potassium channels, particularly those channelsresponsible for the M-current. As used herein, the term “M-current,”“channels responsible for the M-current” and the like, refers to aslowly activating, non-inactivating, slowly deactivating voltage-gatedK+ channel. M-current is active at voltages close to the threshold foraction potential generation in a wide variety of neuronal cells, andthus, is an important regulator of neuronal excitability.

[0049] Recently, members of the voltage-dependent potassium channelfamily were shown to be directly involved in diseases of the central orperipheral nervous system. The benzanilides provided herein are nowshown to act as potassium channel openers, particularly for KCNQ2 andKCNQ3, KCNQ4, KCNQ5 and KCNQ6 as well as the heteromultimer channelssuch as KCNQ2/3, KCNQ3/5 or the M-current.

Description of the Embodiments

[0050] I. Modulators of Voltage-Dependent Potassium Channels

[0051] In view of the above surprising discovery, the present inventionprovides in one aspect, compounds according to Formula I:

[0052] in which the symbol Ar¹ represents a member selected from thegroup consisting of aryl, substituted aryl, heteroaryl and substitutedheteroaryl. The letter X represents a member selected from the groupconsisting of O, S and N—R¹, in which R¹ is H, (C₁-C₈)alkyl, substituted(C₁-C₈)alkyl, heteroalkyl, substituted heteroalkyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, aryl(C₁-C₄)alkyl, substitutedaryl(C₁-C₄)alkyl, CN, —C(O)R², —OR³, —C(O)NR³R⁴, or —S(O)₂NR³R⁴. Thesymbol R² represents a member selected from the group consisting of(C₁-C₈)alkyl, substituted (C₁-C₈)alkyl, cycloalkyl, substitutedcycloalkyl, heteroalkyl, substituted heteroalkyl, heterocyclyl,substituted heterocyclyl, substituted aryl, heteroaryl, substitutedheteroaryl, aryl(C₁-C₄)alkyl and substituted aryl(C₁-C₄)alkyl. R³ and R⁴are each members independently selected from the group consisting ofhydrogen, (C₁-C₈)alkyl, substituted (C₁-C₈)alkyl, cycloalkyl,substituted cycloalkyl, heteroalkyl, substituted heteroalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, aryl(C₁-C₄)alkyl and substitutedaryl(C₁-C₄)alkyl. Alternatively, R³ and R⁴ can be combined with thenitrogen to which each is attached to form a 5-, 6- or 7-membered ring,optionally having additional heteroatoms at the ring vertices. Theletter Y represents a member selected from the group consisting ofhalogen, C₁-C₄ alkyl, C₁-C₄ substituted alkyl, —OCH₃ and —OCF₃.

[0053] In one group of preferred embodiments, Ar¹ is selected fromphenyl, substituted phenyl, indolyl, substituted indolyl, benzofuranyl,substituted benzofuranyl, furanyl, substituted furanyl, thienyl,substituted thienyl, isothiazolyl, substituted isothiazolyl, pyrazolylor substituted pyrazolyl. Still further preferred are those embodimentsin which Ar¹ is substituted phenyl, substituted or unsubstituted2-indolyl and substituted or unsubstituted 2-thienyl. In yet anothergroup of preferred embodiments, X is O.

[0054] In those preferred embodiments, in which Ar¹ is substituted, theAr¹ substituents are preferably halogen, alkyl, halo(C₁-C₄)alkyl,(C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, nitro, cyano, —N R⁷C(O) R⁸, —N R⁷R⁸,phenyl and/or substituted phenyl. The symbols R⁷ and R⁸ independentlyrepresent hydrogen, (C₁-C₈)alkyl, substituted (C₁-C₈)alkyl, cycloalkyl,substituted cycloalkyl, heteroalkyl, substituted heteroalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, aryl(C₁-C₄)alkyl and substitutedaryl(C₁-C₄)alkyl. Alternatively, R⁷ and R⁸ are combined with thenitrogen to which it is attached to form a 5-, 6- or 7-membered ring,optionally having additional heteroatoms at the ring vertices.

[0055] In a further aspect, the present invention provides compoundshaving a structure according to Formula III:

[0056] In Formula III, the symbols Y¹ and Y² independently represent H,methyl, methoxy, trifluoromethoxy, —CF₃ or halo, with the proviso thatboth Y¹ and Y² are not H. The symbols V and X independently represent H,halo, substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, NO₂, CN, CF₃, C(O)NR¹¹R¹² and C(O)R¹³.The symbols R¹¹, R¹² and R¹³ independently represent substituted orunsubstituted lower alkyl, substituted or unsubstituted lowerheteroalkyl, substituted or unsubstituted carbocycle, substituted orunsubstituted heterocycle, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. R¹¹ and R¹² can be joined intoa ring. Q and W independently represent —(CR²R³)_(t)—(CH₂)_(n)—,—CH₂)_(n)—(CR²R³)_(t), —C(R⁴)═C(R⁵)—, or —C≡C— wherein R² and R³ areindependently H, F, substituted or unsubstituted lower alkyl orsubstituted or unsubstituted lower heteroalkyl, wherein R² and R³ areoptionally joined to form a cyclic structure which is a member selectedfrom the group consisting of cycloalkyls and heterocycles, or R² and R³together with the carbon to which they are attached form carbonyl (i.e.,—C(O)—). Q can optionally be a bond between the phenyl ring and Z, and Wcan optionally be a bond between Z and R¹. The symbol Z represents —O—,—S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—, —C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—,—N(R⁴)C(O)O—, —(CR²R³)—, and —SO₂N(R⁴)—, wherein R⁴ and R⁵ are membersindependently selected from the group consisting of H, substituted orunsubstituted lower alkyl, substituted or unsubstituted lowerheteroalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. R¹ is optionally joined together with either Xor R⁴ to form a substituted or unsubstituted heterocycle. The symbol mrepresents an integer from 0 to 2, inclusive; n represents and integerfrom 0 and 3, inclusive; and t represents an integer from 0 to 2,inclusive.

[0057] In yet a further group of preferred embodiments, the compounds ofthe invention have a structure according to Formula IV:

[0058] In Formula IV, R—W—Z—Q— is R⁶, and R⁶ is selected from H,halogen, substituted or unsubstituted alkyl, halo(C₁-C₄)alkyl, nitro,cyano, substituted or unsubstituted phenyl, R⁹O—; R⁹S—; R⁹NH—; R⁹NH—;R⁹NHS(O)₂—; R⁹S(O)₂—, with the proviso that both X and R⁶ are not H. Thesymbol R⁹ represents a member selected from aryl, and alkylaryl; whenthere is more than one R⁹ group per molecule, each R⁹ group isindependently selected. The symbol Y represents a member selected fromhalogen, C₁-C₄ alkyl, —OCH₃, and —OCF₃.

[0059] In a preferred embodiment, the compounds according to Formula IV,include an arylalkyl group at R⁹ in which the aryl component of thearylalkyl group is unsubstituted aryl, or an aryl group that issubstituted by from 1 to 3 halogen moieties. More preferably, the alkylcomponent of the arylalkyl group is a C₁-C₄ alkyl group.

[0060] In another preferred embodiment, the arylalkyl group according toR⁹ is a substituted or unsubstituted heteroarylalkyl group. Preferredheteroarylalkyl groups include (C₁-C₄)alkylpyridyl groups.

[0061] Certain combinations of the above preferred embodiments form agroup of particularly preferred compounds. Accordingly, representativepreferred compounds of the present invention are set forth in FIG. 1,appended hereto.

[0062] Also within the scope of the present invention are compounds ofthe invention that function as poly- or multi-valent species, including,for example, species such as dimers, trimers, tetramers and higherhomologs of the compounds of the invention or reactive analoguesthereof. The poly- and multi-valent species can be assembled from asingle species or more than one species of the invention. For example, adimeric construct can be “homo-dimeric” or “heterodimeric.” Moreover,poly- and multi-valent constructs in which a compound of the inventionor reactive analogues thereof are attached to an oligomeric or polymericframework (e.g., polylysine, dextran, hydroxyethyl starch and the like)are within the scope of the present invention. The framework ispreferably polyfunctional (i.e. having an array of reactive sites forattaching compounds of the invention). Moreover, the framework can bederivatized with a single species of the invention or more than onespecies of the invention.

[0063] Moreover, the present invention includes compounds within themotif set forth in Formulae I-IV, which are functionalized to affordcompounds having a water-solubility that is enhanced relative toanalogous compounds that are not similarly functionalized. Methods ofenhancing the water-solubility of organic compounds is known in the art.Such methods include, but are not limited to, functionalizing an organicnucleus with a permanently charged moiety, e.g., quaternary ammonium, ora group that is charged at a physiologically relevant pH, e.g.carboxylic acid, amine. Other methods include, appending to the organicnucleus hydroxyl- or amine-containing groups, e.g. alcohols, polyols,polyethers, and the like. Representative examples include, but are notlimited to, polylysine, polyethyleneimine, poly(ethyleneglycol) andpoly(propyleneglycol). Suitable functionalization chemistries andstrategies for these compounds are known in the art. See, for example,Dunn, R.L., et al., Eds. POLYMERIC DRUGS AND DRUG DELIVERY SYSTEMS, ACSSymposium Series Vol. 469, American Chemical Society, Washington, D.C.1991.

[0064] Preparation of Potassium Channel Openers

[0065] Compounds of the present invention can be prepared using readilyavailable starting materials or known intermediates. Briefly, thesynthesis of N-aryl benzamides involves formation of a single amide bondfrom a “carbonyl component” (e.g., a carboxylic acid, carboxylic acidchloride, ester or an activated form of a carboxylic acid, for example,a symmetrical or mixed anhydride) and an “amine component” (e.g., ananiline, aniline derivative, amino heterocycle, and the like). Generaland specific procedures for the preparation of the present compounds areprovided in the examples below.

[0066] Other compounds of the present invention can be prepared usingstandard procedures as outlined in Scheme 1 below. In this scheme, anN-phenyl benzamide (i, wherein D¹ and D² represent substituents,including multiple substituents on the aryl groups) can be treated withreagents such as Lawessons's reagent to provide the thioamides, ii.Alkylation of ii, with, for example, methyl iodide produces iii whichcan be converted to target structures iv, v and vi. Thus, treatment ofiii with sodium hydride (or another suitable base) and sulfamideprovides the sulfamoylimino derivative, iv. Similarly, treatment of iiiwith sodium hydride or another base, followed by cyanamide provides v.Conversion of v to vi can be accomplished with HCI.

[0067] One of skill in the art will recognize that other compounds ofthe present invention can be prepared from intermediates such as iii.For example, treatment of iii with a primary or secondary amine willprovide amidine derivatives that are useful as described or they can befurther derivatized.

[0068] Other methods of preparing the benzanilides of the invention willbe apparent to, and are readily accessible by those of skill in the art.

[0069] Methods for preparing dimers, trimers and higher homologs ofsmall organic molecules, such as those of the present invention, as wellas methods of finctionalizing a polyfunctional framework molecule arewell known to those of skill in the art. For example, an aromatic amineof the invention is converted to the corresponding isothiocyanate by theaction of thiophosgene. The resulting isothiocyanate is coupled to anamine of the invention, thereby forming either a homo- or hetero-dimericspecies. Alternatively, the isothiocyanate is coupled with anamine-containing backbone, such as polylysine, thereby forming aconjugate between a polyvalent framework and a compound of theinvention. If it is desired to prepare a hetereofuntionalized polyvalentspecies, the polylysine is underlabeled with the first isothiocyanateand subsequently labeled with one or more different isothiocyanates.Alternatively, a mixture of isothiocyanates is added to the backbone.Purification proceeds by, for example, size exclusion chromatography,dialysis, nanofiltration and the like.

[0070] II. ASSAYS FOR MODULATORS OF KCNQ CHANNELS

[0071] The present invention also provides assays for determining theability of a compound of the invention to open a potassium ion channel.For simplicity, portions of the following discussion focuses on KCNQ2 asa representative example, however, the discussion is equally applicableto other potassium ion channels.

[0072] KCNQ monomers as well as KCNQ alleles and polymorphic variantsare subunits of potassium channels. The activity of a potassium channelcomprising KCNQ subunits can be assessed using a variety of in vitro andin vivo assays, e.g., measuring current, measuring membrane potential,measuring ion flux, e.g., potassium or rubidium, measuring potassiumconcentration, measuring second messengers and transcription levels,using potassium-dependent yeast growth assays, and using e.g.,voltage-sensitive dyes, radioactive tracers, and patch-clampelectrophysiology.

[0073] Furthermore, such assays can be used to test for inhibitors andactivators of channels comprising KCNQ. Such modulators of a potassiumchannel are useful for treating various disorders involving potassiumchannels discussed herein and known to those of skill in the art.Exemplary disorders include, but are not limited to, central andperipheral nervous system disorders (e.g., migraine, ataxia, Parkinson'sdisease, bipolar disorders, spasticity, mood disorders, brain tumors,psychotic disorders, myokymia, seizures, epilepsy, hearing and visionloss, Alzheimer's disease, age-related memory loss, learningdeficiencies, and motor neuron diseases, and can also be used asneuroprotective agents (e.g., to prevent stroke and the like). Suchmodulators are also useful for investigation of the channel diversityprovided by KCNQ and the regulation/modulation of potassium channelactivity provided by KCNQ.

[0074] Modulators of the potassium channels are tested usingbiologically active KCNQ, either recombinant or naturally occurring, orby using native cells, like cells from the nervous system expressing theM-current. KCNQ can be isolated, co-expressed or expressed in a cell, orexpressed in a membrane derived from a cell. In such assays, KCNQ2 isexpressed alone to form a homomeric potassium channel or is co-expressedwith a second subunit (e.g., another KCNQ family member, preferablyKCNQ3) so as to form a heteromeric potassium channel. Modulation istested using one of the in vitro or in vivo assays described above.Samples or assays that are treated with a potential potassium channelinhibitor or activator are compared to control samples without the testcompound, to examine the extent of modulation. Control samples(untreated with activators or inhibitors) are assigned a relativepotassium channel activity value of 100. Activation of channelscomprising KCNQ2 is achieved when the potassium channel activity valuerelative to the control is 110%, more preferably 130%, more preferably170% higher. Compounds that increase the flux of ions will cause adetectable increase in the ion current density by increasing theprobability of a channel comprising KCNQ2 being open, by decreasing theprobability of it being closed, by increasing conductance through thechannel, and increasing the number or expression of channels.

[0075] Changes in ion flux may be assessed by determining changes inpolarization (i.e., electrical potential) of the cell or membraneexpressing the potassium channel comprising, for example, KCNQ2, KCNQ2/3or the M-current. A preferred means to determine changes in cellularpolarization is by measuring changes in current or voltage with thevoltage-clarnp and patch-clamp techniques, using the “cell-attached”mode, the “inside-out” mode, the “outside-out” mode, the “perforatedcell” mode, the “one or two electrode” mode, or the “whole cell” mode(see, e.g., Ackerman et al., New Engl. J Med. 336:1575-1595 (1997)).Whole cell currents are conveniently determined using the standardmethodology (see, e.g., Hamil et al., Pflugers. Archiv. 391:85 (1981).Other known assays include: radiolabeled rubidium flux assays andfluorescence assays using voltage-sensitive dyes (see, e.g.,Vestergarrd-Bogind et al., J Membrane Biol. 88:67-75 (1988); Daniel etal., J. Pharmacol. Meth. 25:185-193 (1991); Holevinsky et al., JMembrane Biology 137:59-70 (1994)). Assays for compounds capable ofinhibiting or increasing potassium flux through the channel proteinscomprising KCNQ2 or heteromultimers of KCNQ subunits can be performed byapplication of the compounds to a bath solution in contact with andcomprising cells having a channel of the present invention (see, e.g.,Blatz et al., Nature 323:718-720 (1986); Park, J Physiol. 481:555-570(1994)). Generally, the compounds to be tested are present in the rangefrom 1 pM to 100 mnM.

[0076] The effects of the test compounds upon the flnction of thechannels can be measured by changes in the electrical currents or ionicflux or by the consequences of changes in currents and flux. Changes inelectrical current or ionic flux are measured by either increases ordecreases in flux of ions such as potassium or rubidium ions. Thecations can be measured in a variety of standard ways. They can bemeasured directly by concentration changes of the ions or indirectly bymembrane potential or by radio-labeling of the ions. Consequences of thetest compound on ion flux can be quite varied. IAccordingly, anysuitable physiological change can be used to assess the influence of atest compound on the channels of this invention. The effects of a testcompound can be measured by a toxin binding assay. When the finctionalconsequences are determined using intact cells or animals, one can alsomeasure a variety of effects such as transmitter release (e.g.,dopamine), hormone release (e.g., insulin), transcriptional changes toboth known and uncharacterized genetic markers (e.g., northern blots),cell volume changes (e.g., in red blood cells), immunoresponses (e.g., Tcell activation), changes in cell metabolism such as cell growth or pHchanges, and changes in intracellular second messengers such as Ca²⁺, orcyclic nucleotides.

[0077] Preferably, the KCNQ2, or other KCNQ channel constituents that isa part of the potassium channel used in the assay will have the sequenceprovided in PCTIUS98/13276 or a conservatively modified variant thereofAlternatively, the KCNQ2 of the assay will be derived from a eukaryote.

[0078] KCNQ2 orthologs will generally confer substantially similarproperties on a channel comprising such KCNQ2, as described above. In apreferred embodiment, the cell placed in contact with a compound that issuspected to be a KCNQ2 homolog is assayed for increasing or decreasingion flux in a eukaryotic cell, e.g., an oocyte of Xenopus (e.g., Xenopuslaevis) or a mammalian cell such as a CHO or HeLa cell. Channels thatare affected by compounds in ways similar to KCNQ2 are consideredhomologs or orthologs of KCNQ2.

[0079] III. PHARMACEUTICAL COMPOSITIONS OF POTASSIUM CHANNEL OPENERS

[0080] In another aspect, the present invention provides pharmaceuticalcompositions comprising a pharmaceutically acceptable excipient and acompound according to Formula II, above.

[0081] Formulation of the Compounds (Compositions)

[0082] The compounds of the present invention can be prepared andadministered in a wide variety of oral, parenteral and topical dosageforms. Thus, the compounds of the present invention can be administeredby injection, that is, intravenously, intramuscularly, intracutaneously,subcutaneously, intraduodenally, or intraperitoneally. Also, thecompounds described herein can be administered by inhalation, forexample, intranasally. Additionally, the compounds of the presentinvention can be administered transdermally. Accordingly, the presentinvention also provides pharmaceutical compositions comprising apharmaceutically acceptable carrier or excipient and either a compoundof Formulae I-lV or a pharmaceutically acceptable salt of a compound ofFormulae I-IV.

[0083] For preparing pharmaceutical compositions from the compounds ofthe present invention, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, pills, capsules, cachets, suppositories, and dispersiblegranules. A solid carrier can be one or more substance, which may alsoact as diluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material.

[0084] In powders, the carrier is a finely divided solid, which is in amixture with the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired.

[0085] The powders and tablets preferably contain from 5% or 10% to 70%of the active compound. Suitable carriers are magnesium carbonate,magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch,gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, alow melting wax, cocoa butter, and the like. The term preparation isintended to include the formulation of the active compound withencapsulating material as a carrier providing a capsule in which theactive component with or without other carriers, is surrounded by acarrier, which is thus in association with it. Similarly, cachets andlozenges are included. Tablets, powders, capsules, pills, cachets, andlozenges can be used as solid dosage forms suitable for oraladministration.

[0086] For preparing suppositories, a low melting wax, such as a mixtureof fatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

[0087] Liquid form preparations include solutions, suspensions, andemulsions, for example, water or water/propylene glycol solutions. Forparenteral injection, liquid preparations can be formulated in solutionin aqueous polyethylene glycol solution.

[0088] Aqueous solutions suitable for oral use can be prepared bydissolving the active component in water and adding suitable colorants,flavors, stabilizers, and thickening agents as desired. Aqueoussuspensions suitable for oral use can be made by dispersing the finelydivided active component in water with viscous material, such as naturalor synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, and other well-known suspending agents.

[0089] Also included are solid form preparations, which are intended tobe converted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

[0090] The pharmaceutical preparation is preferably in unit dosage form.In such form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

[0091] The quantity of active component in a unit dose preparation maybe varied or adjusted from 0.1 mg to 10000 mg, more typically 1.0 mg to1000 mg, most typically 10 mg to 500 mg, according to the particularapplication and the potency of the active component. The compositioncan, if desired, also contain other compatible therapeutic agents.

[0092] IV. METHODS FOR INCREASING ION FLOW IN VOLTAGE-DEPENDENTPOTASSIUM CHANNELS

[0093] In yet another aspect, the present invention provides methods forincreasing ion flow through voltage dependent potassium channels in acell, comprising contacting a cell containing the target ion channelswith a compound of Formulae I-IV, above.

[0094] The methods provided in this aspect of the invention are usefulfor the diagnosis of conditions that can be treated by modulating ionflux through voltage-dependent potassium channels, or for determining ifa patient will be responsive to therapeutic agents which act by openingpotassium channels. In particular, a patient's cell sample can beobtained and contacted with a compound of Formula I-IV and the ion fluxcan be measured relative to a cells ion flux in the absence of acompound of Formula I-IV. An increase in ion flux will typicallyindicate that the patient will be responsive to a therapeutic regimen ofion channel openers.

[0095] V. METHODS FOR TREATING CONDITIONS MEDIATED BY VOLTAGE DEPENDENTPOTASSIUM CHANNELS

[0096] In still another aspect, the present invention provides a methodfor the treatment of diseases or conditions mediated, at least in part,by voltage-dependent potassium channels. In this method, a subjectsuffering from such a condition or disease is administered an effectiveamount of a compound of Formulae I-IV.

[0097] The compounds provided herein are usefuil as potassium channelopeners and find therapeutic utility via modulation of voltage-dependentpotassium channels in the treatment of diseases or conditions. Thepotassium channels that are typically opened are described herein asvoltage-dependent potassium channels such as the KCNQ potassiumchannels. As noted above, these channels may include homomultimers andheteromultimers of KCNQ2, KCNQ3, KCNQ4, KCNQ5 and KCNQ6. Aheteromultimer of two proteins, e.g., KCNQ2 and KCNQ3 is referred to as,for example, KCNQ2/3. The conditions that can be treated with thecompounds and compositions of the present invention may include, but arenot limited to, central or peripheral nervous system disorders (e.g.,migraine, ataxia, Parkinson's disease, bipolar disorders, spasticity,mood disorders, brain tumors, psychotic disorders, myokymia, seizures,epilepsy, hearing and vision loss, Alzheimer's disease, age-relatedmemory loss, learning deficiencies, and motor neuron diseases, and asneuroprotective agents (e.g., to prevent stroke and the like)).

[0098] In therapeutic use for the treatment of epilepsy or otherneurological conditions, the compounds utilized in the pharmaceuticalmethod of the invention are administered at the initial dosage of about0.001 mg/kg to about 1000 mg/kg daily. A daily dose range of about 0.1mg/kg to about 100 mg/kg is more typical. The dosages, however, may bevaried depending upon the requirements of the patient, the severity ofthe condition being treated, and the compound being employed.Determination of the proper dosage for a particular situation is withinthe skill of the practitioner. Generally, treatment is initiated withsmaller dosages, which are less than the optimum dose of the compound.Thereafter, the dosage is increased by small increments until theoptimum effect under circumstances is reached. For convenience, thetotal daily dosage may be divided and administered in portions duringthe day, if desired.

[0099] The materials, methods and devices of the present invention arefurther illustrated by the examples, which follow. These examples areoffered to illustrate, but not to limit the claimed invention.

EXAMPLES

[0100] Example 1 sets forth representative methods of preparing2-substituted -5-aminopyridines of use in preparing the compounds of theinvention. The representative methods include the reduction ofnitropyridines, rearrangement of nicotinic acids, and the displacementreactions of 2-halopyridines.

[0101] Examples 2 and 3 set forth a representative methods of preparingthe benzanilides of the invention. Example 2 provides a method ofpreparing a benzanilide form an acid chloride. Example 3 provides amethod of preparing a benzanilide from a carboxylic acid by generatingthe acid chloride in situ.

[0102] Example 4, 5, 6, and 7 set forth methods of elaborating thebenzanilide nucleus. Example 4 provides a method of preparing 4-aminosubstituted benzanilides via a nucleophilic displacement. Example 5provides a method for reducing an aromatic nitro group to thecorresponding amine. Example 6 provides a method for preparing hydroxylamine compounds. Example 7 provides a method for preparing sulfonamides.Example 8 sets forth the characterization of the compounds prepared bythe methods of Examples 1-7.

[0103] Examples 8 and 9 set forth the preparation of a number ofrepresentative compounds of the invention in which the benzanilidenucleus is elaborated by selected groups. Example 10 sets forth theresults of the physical characterization of the compounds prepared byExamples 8 and 9. Example 11 sets forth an assay usefiul in theevaluation of the activity towards potassium ion channels, e.g., KCNQ2,of selected compounds of the invention.

[0104] General

[0105] In the examples below, unless otherwise stated, temperatures aregiven in degrees Celsius (° C); operations were carried out at room orambient temperature, “rt,” or “RT,” (typically a range of from about18-25° C.; evaporation of solvent was carried out using a rotaryevaporator under reduced pressure (typically, 4.5-30 mmnHg) with a bathtemperature of up to 60° C.; the course of reactions was typicallyfollowed by TLC and reaction times are provided for illustration only;melting points are uncorrected; products exhibited satisfactory ¹H-NMRand/or microanalytical data; yields are provided for illustration only;and the following conventional abbreviations are also used: mp (meltingpoint), L (liter(s)), mL (milliliters), mmol (millimoles), g (grams), mg(milligrams), min (minutes), and h (hours).

[0106] General Experimental.

[0107] Unless otherwise specified, all solvents (HPLC grade) andreagents were purchased from suppliers and used without furtherpurification. Reactions were conducted under a blanket of argon unlessotherwise stated. Analytical thin layer chromatography (tlc) wasperformed on Whatman Inc. 60 silica gel plates (0.25 mm thickness).Compounds were visualized under UV lamp (254 nM) or by developing withKMnO4/KOH, ninhydrin or Hanessians solution. Flash chromatography wasdone using silica gel from Selcetro Scientific (particle size 32-63). ¹HNMR, ¹⁹F NMR and ¹³C NMR spectra were recorded on a Varian 300 machineat 300 MHz, 282 MHz and 75.7 MHz, respectively. Melting points wererecorded on a Electrothermal IA9100 apparatus and were uncorrected.

EXAMPLE 1

[0108] Preparation of 2-substituted-5-aminopyridines

[0109] 1.1 Reduction of nitropyridines

[0110] Referring to Scheme 2, the desired aminopyridines (II) areprepared by reducing the corresponding nitropyridines (I). One skilledin the art will recognize that there are several methods to accomplishstep 1. Tin chloride in DMF, hydrogenation using catalytic palladium andsodium borohydride in the presence of catalytic nickel chloride areknown methods.

[0111] 1.1a Synthesis of 5-amino-2-bromopyridine

[0112] Tin (II) chloride hydrate (0.78 g, 3.5 mmol) was added to astirring solution of 5-nitro-2-bromopyridine (0.24 g, 1.2 mmol) in DMF(5 mL) at RT. After 2 h, 6 N NaOH (2 mL) was added and the suspensionwas stirred vigorously for 10 min. The organics were extracted withdiethyl ether (2×10 mL), washed with brine (2×10 mL) and dried (Na₂SO₄).The filtered solution was then concentrated under reduced pressure toafford the desired product as a yellow oil (0.178 g, 86%), which wasused without further purification.

[0113] 1.2 Rearrangement of nicotinic acids

[0114] Rearrangement of the corresponding nicotinic acids (III) (Scheme3) using a modified Schmidt reaction, followed by deprotection of theaniline group generated the desired aminopyridines (IV) as thecorresponding TFA salts.

[0115] 1.2a Synthesis of 5-amino-2-methylpvridine (TFA salt)

[0116] A solution of diphenylphosphorylazide (430 μL, 2 mmol),triethylamine (278 μL, 2 mmol) and 6-methyl-nicotinic acid (274 mg, 2mmol) in t-butanol (30 mL) was heated at reflux for 4 h. The solutionwas cooled to RT and poured into water (50 mL). The organics wereextracted with ether (3 x 20 mL), washed with brine (2 x 10 mL) anddried (Na₂SO₄). Column chromatography (1:1 hexane/ethyl acetate) of theboc-protected aminopyridine gave the intermediate as a white solid (156mg, 38%).

[0117] The desired 5-amino-2-methylpyridine-TFA salt was generated insitu by stirring in a 20% TFA/DCM solution (2 mL) for 4 h. The solutionwas concentrated under reduced pressure to afford a semi-solid, whichwas used without further purification. 1.2b Synthesis of5-amino-2-(trifluoromethyl)pyridine (TFA salt)

[0118] A solution of diphenylphosphorylazide (644 μL, 3 mnmol),triethylamine (417 μL, 3 rnmol) and 6-(trifluoromethyl)-nicotinic acid(573 mg, 3 mmol) in t-butanol (50 mL) was heated at reflux for 4 h, thencooled to RT and poured into water (50 mL). The organics were extractedwith ether (3×20 mL), washed with brine (2×10 mL) and dried (Na₂SO₄).Column chromatography (1:1 hexane/ethyl acetate) of the boc-protectedaniline gave the intermediate as a white solid (389 mg, 50%).

[0119] The desired 5-amino-2-methylpyridine-TFA salt was generated insitu by stirring in a 20% TFA/DCM solution (2 mL) for 4 h. The solutionwas concentrated under reduced pressure to afford a semi-solid, whichwas used without further purification.

[0120] 1.3: Displacement of 2-halopyridines.

[0121] Several aminopyridines, which are not readily accessible via themethods outlined in schemes 1 or 2, may be synthesized via nucleophilicdisplacement of 2-chloropyridines as depicted in Scheme 4.

[0122] 1. 3a Synthesis of 5-amino-2-fluoropyridine

[0123] A mixture of 5-nitro-2-chloropyridine (2.0 g, 12.6 mrnol) andanhydrous potassium fluoride (2.2 g, 38 mmol) in a combination ofsulfalone (6 nmL) and benzene (4 mL) was stirred at RT for 20 min. Thebenzene was then removed by distillation. The resulting mixture washeated at 150° C. for 12 h. The mixture was cooled to RT whereupon water(60 mL) was added. The desired product was separated from the solutionvia steam distillation. Extraction of the distillate with diethyl ether(2×10 mL) followed by drying (Na₂SO₄) and concentration gave5-nitro-2-fluoropyridine as a water white oil (1.3 g, 73%).

[0124] 10% Palladium on charcoal (20 mg, cat) was added to a stirringsolution of 5-nitro-2-fluoropyridine (100 mg, 0.7 mmol) indichloromethane (3 mL) at RT. 1 atmosphere of hydrogen gas was thenapplied to the solution and the mixture was stirred at RT for 1 h. Themixture was passed through a short plug of celite and the resultingsolution, containing the desired 5-amino-2-fluoropyridine, was usedwithout further purification.

EXAMPLE 2

[0125] 2.1 Preparation of Benzanilides from Acid Chlorides

[0126] Benzanilides (e.g., VIII) were prepared by reacting acidchlorides (e.g., VII) with aminopyridines (e.g., II, IV and VI) as shownin Scheme 5. The reaction was typically conducted in the presence of atertiary amine base such as triethylamine in an organic solvent such asdichloromethane or tetrahydrofuran, and at room temperature.

[0127] 2.2 General Experimental for Scheme 5

[0128] A solution of acid chloride (VII) (1 mmol) in a dry solvent(e.g., acetonitrile, THF, DCM) (3 mL) was added dropwise to a stirringsolution of aminopyridine (II or VI) (1 mmol) and N,N-diisopropylethylamine (1.2 mmol) in a dry solvent (e.g., acetonitrile,THF, DCM) (5 mL) at RT. The resulting solution was stirred for anadditional 1 h. If TLC analysis indicated presence of starting anilinethe solution was heated at 55° C. for another 1 h. After cooling to roomtemperature ethyl acetate (10 mL) was added and the solution was washedwith water (2×10 mL) and dried (Na₂SO₄). The solvent was removed underreduced pressure and the crude material was purified by columnchromatography (hexanes/ethyl acetate) or by crystallization(hexane/dichloromethane). The products were typically white solids(50-98%). Compounds prepared via this procedure include, 1-15, 25, 27,29 and 42.

[0129] 2.2a Preparation ofN—[2-chloro-5-pyridyl]-3-(trifluoromethyl)benzamide (3)

[0130] To a stirring solution of 5-amino-2-chloropyridine (129 mg, 1mmol) and N,N-diisopropylethylamine (209 μL, 1.2 mmol) in dryacetonitrile (5 mL) was added 3-(trifluoromethyl)benzoyl chloride (151pL, 1 mmol). The resulting solution was heated at 55° C. for 3 h. Aftercooling to room temperature ethyl acetate (10 mL) was added and thesolution was washed with water (2×10 mL) and dried (Na₂SO₄). The solventwas removed under reduced pressure and the crude material was purifiedby column chromatography (4:1, hexane/ethyl acetate) to afford thedesired product as a white solid (284 mg, 94%).

EXAMPLE 3

[0131] Preparation of Benzanilides from Acids

[0132] Benzanilides (VIII) may be also be prepared from acids (IX) byinitially converting them to their acid chlorides (VII). Acids (IX) weretreated with oxalyl chloride in the presence of catalytic N,N-dimethylformamide in an organic solvent such as dichloromethane ortetrahydrofuran preferably at 0° C. The acid chloride, generated in situwas then reacted with aminopyridines (II, IV or VI) in the presence of atertiary amine base such as triethylamine in an organic solvent such asdichloromethane or tetrahydrofuran. The reactions were typicallyperformed at RT.

[0133] 3.1 General Experimental for Scheme 6

[0134] Oxalyl chloride (1.05 mmol) was added dropwise to a stirringsuspension of acid (IX) (1 mmol) and DMF (0.1 mmol) in dry DCM (5 mL) at0° C. Once addition was complete the reaction was allowed to warn to RTand stirred for a further 45 min whereupon the reaction was a clearsolution. This solution was added dropwise to a stirring solution ofaminopyridine (II or VI) (0.95 mmol) and N,N-diisopropylethylamine (2.2mmol) in DCM (5 mL) at RT. After 30 min the organics were washed withaqueous 1 N NaOH (10 mL), brine (10 mL) and dried (Na₂SO₄). The filteredsolution was concentrated under reduced pressure and the crude productwas purified by column chromatography (hexanes/ethyl acetate) or bycrystallization (hexane/dichloromethane). Compounds prepared via thisprocedure include, compounds 16-24, 26, 28 and 37.

EXAMPLE 4

[0135] Preparation of 4-Amino Substituted Benzamides via NucleophilicDisplacement

[0136] Aryl fluorides (X) (Scheme 7) possessing strongly electronwithdrawing groups in either the ortho or para positions were displacedwith primary or secondary amines under elevated temperatures in a polarorganic solvent such as DMSO or NMP to yield compounds of the formula(XI).

[0137] 4.1 General Experimental for Scheme 7

[0138] A solution of amine (1.1 mmol) and (X) (1 mmol) in either dry NMPor DMSO (3 mL) was heated at 120° C. for 12 h. After cooling to RT,water (10 mL) and ethyl acetate (10 mL) were added. After vortexing forseveral minutes the organic layer was removed and dried (Na₂SO₄). Thesolvent was removed under reduced pressure and the residue purified bycolumn chromatography (hexanes/ethyl acetate or acetone/chloroform). Theproducts (XI) were obtained as white solids (30-50%). Compounds preparedvia this procedure include, compounds 30-34.

EXAMPLE 5

[0139] Reduction of Aromatic Nitro to Amine

[0140] Scheme 8 outlines a general synthetic route to compounds offormulae (XIII) and (XIV).

[0141] 5.1 General experimental for Scheme 8

[0142] Compounds of structure (XII) were prepared using either generalprocedure B or C. Tin (II) chloride hydrate (22 mmol) was added to astirring solution of (XII) in DMF (20 mL) at RT. After 14 h, 6 N NaOH (6mL) was added and the suspension was stirred vigorously for 10 min. Theorganics were extracted with ethyl acetate (2×20 mL), washed with brine(2×10 mL) and dried (Na₂SO₄). The filtered solutions were concentratedunder reduced pressure and the crude products were purified by columnchromatography (hexanes/ethyl acetate; 1:2) to afford the desiredintermediates (XIII) as a white solids (60-90%).

[0143] The intermediates (XIII) (0.2 mmol) were coupled with either acidchlorides (VI) or acids (0.2 mmol) using the methods described ingeneral procedures A and B. The desired products (XIV) were obtained astan solids (20-60%). Compounds prepared via this procedure include,compound 35.

EXAMPLE 6

[0144] Preparation of 4-Hydroxyamino Compounds (XV).

[0145] Hydroxyamines (XV) were prepared according to the synthetic routeoutlined in Scheme 9.

[0146] 6.1 General Experimental for Scheme 9

[0147] Tin (II) chloride hydrate (3 mmol) was added to a stirringsolution of nitrobenzamide (XII) (1 mmol) in DMF (5 mL) at RT. After 3h, 6 N NaOH (6 mL) was added and the suspension was stirred vigorouslyfor 10 min. The organics were extracted with ethyl acetate (2×20 mL),washed with brine (2×10 mL) and dried (Na₂SO₄). The filtered solutionwas then concentrated under reduced pressure to afford the crudeproduct. Purification by column chromatography (ethyl acetate) gave thedesired products (XV) as beige solids (50-70%).

EXAMPLE 7

[0148] Preparation of Sulfonamides

[0149] Sulfonamides (e.g., XVIII) were prepared using the chemistryoutlined in Scheme 10. Intermediate (XVII) were generated by coupling anaminopyridine (II or IV) with an activated form of (XVI). Subsequentcoupling of the sulfonyl group with an amine generated the desiredsulfonamides (XVIII)

[0150] 7.1 General Experimental for Scheme 10

[0151] Oxalyl chloride (175 μL, 2 mmol) was added dropwise to a stirringsolution of (XVI) (440 mg, 2 mmol) and DMF (20 μL, cat) in THF (8 mL) at0° C. After addition was complete the reaction was allowed to warm toRT. After 30 min the reaction was cooled back to 0° C. whereupon asolution of (II or IV) (1.9 mmol) and N, N-diisopropylethylamine (700μL, 4 mmol) in THF (2.3 mL) was added. After stirring for 30 min at RT,this solution of (XVII) (0.18 M) was used directly without furthermanipulation.

[0152] A solution of (XVII) (5.5 mL, 1 mmol) in THF was added to astirring solution of amine (1 mmol) and N,N-diisopropylethylamine (3mmol) in THF (2 mL) at RT. After 1 h, water (10 mL) and ethyl acetate(10 mL) were added. The organic layer was separated, washed with water(5 mL), aqueous 1N NaOH (5 mL), aqueous 1N HCl (5 mL) and then dried(Na₂SO₄). The solvent was removed under reduced pressure and the residuepurified by column chromatography (hexanes/ethyl acetate; 1:2). Theproducts were obtained as white solids (60-80%). Compounds prepared viathis procedure include, e.g., compounds 36 and 40.

EXAMPLE 8

[0153] Example 8 sets forth the results of the characterization ofrepresentative compounds prepared by the methods of Examples 1 through7. The compounds were characterized using a combination of meltingpoint, ¹H NMR and mass spectrometry. The results of the characterizationare presented below. The structures for the compounds set forth beloware provided in FIG. 1.

[0154] 3,4-Dichloro-N-pyridin-3-yl-benzamide (1)

[0155] mp 165-166° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.58 (1 H, brs), 8.87(1 H, d, J═2.3 Hz), 8.30 (1 H, dd, J═4.7, 1.4 Hz), 8.19 (1 H, d, J═1.9Hz), 8.16-8.12 (1 H, m), 7.91 (1 H, dd, J═8.4, 2.1 Hz), 7.80 (1 H, d,J═8.5 Hz) and 7.38 (1 H, dd, J═8.4, 4.7 Hz); MS (ESI) m/z: 266.9 [M+H]⁺.

[0156] 3,4-Dichloro-N-(6-chloro-pyridin-3-yl)-benzamide (2)

[0157] mp 188-189° C.;¹H NMR (300 MHz, CDCI₃) δ7.36 (1 H,d, J═8.7 Hz),7.58 (1 H, d, J═9.3 Hz), 7.70 (1 H, dd, J═9.4, 2.0 Hz), 7.90 (1 H, brs),7.96 (1 H, d, J═1.9 Hz), 8.24 (1 H, dd, J═8.7, 2.8 Hz) and 8.48 (1 H, d,J═2.8 Hz); ¹³C NMR (75 MHz, DMSO-d₆) δ124.7, 128.6, 130.2, 131.4, 131.9,134.8, 135.4, 142.0, 144.9 and 164.0; MS (ESI) m/z: 301.1 [M+H]+.

[0158] N-(6-Chloro-pyridin-3-yl)-3-trifluoromethyl-benzamide (3)

[0159] mp 139-140° C.; ¹H NMR (300 MHz, CDCI₃) Δ7.35 (1 H, d, J═8.7 Hz),7.63 (1 H, t, J═7.8 Hz), 7.82 (1 H, d, J═7.8 Hz), 8.06 (1 H, d, J═7.8Hz), 8.11 (1 H, brs), 8.25 (1 H, dd, J═8.7, 2.9 Hz), 8.36 (1 H, s) and8.50 (1 H, d, J═2.6 Hz); ¹⁹F NMR (282 MHz, CDCI₃) Δ−63.6; ¹³C NMR (75MHz, DMSO-d₆) δ124.2, 124.2, 124.6, 129.0, 129.7, 130.6, 130.9, 133.7,134.6, 141.2 and 164.8; MS (ESI) m/z: 301.2 [M+H]⁺.

[0160] N-(6-Chloro-pyridin-3-yl)-3,4-difluoro-benzamide (4)

[0161] mp; 164° C.;¹H NMR (300 MHz, DMSO-d₆) δ7.51 (1 H, d, J═8.7 Hz),7.59-7.68 (1 H, m), 7.84-7.88 (1 H, m), 8.03 (1 H, ddd, J═11.3, 7.9, 2.1Hz), 8.19-8.23 (1 H, m), 8.25 (1 H, d, J═2.0 Hz) and 10.64 (1 H, s); ¹⁹FNMR (282 MHz, DMSO—-d₆) δ-112.9 (m), -115.2 (m); ¹³C NMR (75 MHz,DMSO-d₆) δ117.9 (dd, J═18.3, 49.8 Hz), 124.7, 125.7 (dd, J═3.4, 6.9 Hz),131.5 (m), 131.9, 135.3, 142.1, 145.0, 149.3 (dd, J═14.7, 201.0 Hz),152.8 (dd, J═12.6, 205.0 Hz), 164.6; MS (ESI) m/z: 269.1 [M+H]⁺.

[0162] 3-Chloro-N-(6-chloro-pyridin-3-yl)-benzamide (5)

[0163] mp 153-154° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.67 (1 H, brs), 8.74(1 H, d, J ═2.6 Hz), 8.22 (1 H, dd, J═8.7, 2.8 Hz), 7.99 (1 H, d, J═1.7Hz), 7.90 (1 H, d, J═7.8 Hz), 7.69 (1 H, d, J═7.1 Hz), 7.58 (1 H, t,J═7.8 Hz) and 7.51 (1 H, d, J═8.7 Hz); MS (ESI) m/z: 267.0 [M+H]⁺.

[0164] Biphenyl-4-carboxylic acid (6-chloro-pyridin-3-yl)-amide (6)

[0165] mp 227-229° C.;¹H NMR (300 MHz, DMSO—d₆) δ10.62 (1 H, brs), 8.81(1 H, d, J═2.3 Hz), 8.26 (1 H, dd, J═8.7, 2.4 Hz), 8.06 (2 H, d, J═8.2Hz), 7.85 (2 H, d, J═8.2 Hz), 7.75 (2 H, d, J═7.5 Hz), 7.53-7.47 (3 H,m) and 7.42 (1 H, q, J═7.1 Hz); MS (ESI) m/z: 309.2 [M+H]⁺.

[0166] 6-Chloro-N-(6-chloro-pyridin-3-yl)-nicotinamide (7)

[0167] mp 228° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.81 (1 H, brs), 8.92 (1H, d, J═2.3 Hz), 8.73 (1 H, d, J═2.6 Hz), 8.32 (1 H, dd, J═8.4, 2.4 Hz),8.19 (1 H, dd, J═8.7, 2.8 Hz), 7.71 (1 H, d, J═8.4 Hz) and 7.52 (1 H, d,J═8.7 Hz); MS (ESI) m/z: 268.1 [M+H]⁺.

[0168] 3,4-Difluoro-N-(6-methyl-pyridin-3-yl)-benzamide (8) ¹H NMR (300MHz, DMSO-d₆) δ10.42 (1 H, brs), 8.73 (1 H, d, J═2.3 Hz), 8.04-7.97 (2H, m), 7.86-7.82 (1 H, m), 7.90 (1 H, dt, J═10.4, 8.4 Hz), 7.23 (1 H, d,J═8.5 Hz) and 2.42 (3 H, s); ¹⁹F NMR (282 MHz, DMSO-d₆) δ-133.1 to-133.3 (1 H, m) and -137.1 (1 H, q, J═10.7 Hz); MS (ESI) m/z: 249.0[M+H]⁺.

[0169] N-(6-Chloro-pyridin-3-yl)-3-fluoro-benzamide (9)

[0170] mp 160° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.63 (1 H, brs), 8.75 (1H, d, J ═2.8 Hz), 8.20 (1 H, dd, J═8.7, 2.8 Hz), 7.79 (1 H, d, J═7.8Hz), 7.75 (1 H, d, J═11.1 Hz), 7.62-7.55 (1 H, m) and 7.53-7.43 (2 H,m); ¹⁹F NMR (282 MHz, DMSO-d₆) δ-112.0 (q, 8.5 Hz); MS (ESI) m/z: 251.0[M+H]⁺.

[0171] N-(6-Chloro-pyridin-3-yl)-3-(trifluoromethyl)-benzamide (10)

[0172] mp 169-170° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.81 (1 H, brs), 8.75(1H, d, J ═2.8 Hz), 8.22 (1 H, dd, J═8.7, 2.8 Hz), 8.13 (1 H, d, J═8.2Hz), 7.91 (2 H, d, J═8.4 Hz) and 7.52 (2 H, d, J═8.7 Hz); ¹⁹F NMR (282MHz, DMSO-d₆) δ-61.4 (s); MS (ESI) m/z: 301.2 [M+H]⁺.

[0173] N-(6-Chloro-pyridin-3-vl)-4-fluoro-3-trifluoromethyl-benzamide(11)

[0174] mp 149-150° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.63 (1 H, brs), 8.76(1 H, d, J ═2.6 Hz), 8.22 (1 H, dd, J═8.7,2.8 Hz), 7.98 (2 H, d,J═8.7Hz), 7.63 (1 H, d,J═8.7 Hz) and 7.52 (1 H, d, J═8.7 Hz); MS (ESI) m/z:319.1 [M+H]⁺.

[0175] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-trifluoromethyl-benzamide(12)

[0176] mp 182° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.78 (1 H, brs), 8.75 (1H, d, J ═2.3 Hz), 8.37-8.32 (2 H, m), 8.22 (1 H, dd, J═8.7,2.6 Hz), 7.73(1 H, m) and 7.54 (1 H, d, J═8.7 Hz);¹⁹F NMR (282 MHz, DMSO-d₆) δ-60.1(3 F, m), −110.7 (m); MS (ESI) m/z: 319.1 [M+H]⁺.

[0177] N-(6-Chloro-pyridin-3-yl)-4-fluoro-benzamide (13)

[0178] mp 163-164° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.58 (1 H, brs), 8.76(1 H, d, J ═2.6 Hz), 8.22 (1 H, dd, J═8.7, 2.6 Hz), 8.04 (1 H, d, J═8.7Hz), 8.02 (1 H, d, J═8.7 Hz), 7.51 (1 H, d, J═8.7 Hz) and 7.39 (2 H, t,J═8.8 Hz); ¹⁹F NMR (282 MHz, DMSO-d₆) δ-107.7 (m); MS (ESI) m/z: 319.0[M+H]⁺.

[0179] N-(6-Chloro-pyridin-3-yl)-4-chloro-benzamide (14)

[0180] mp 197-199° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.63 (1 H, brs), 8.76(1 H, d, J═2.6 Hz), 8.22 (1 H, dd, J═8.7, 2.8 Hz), 7.98 (2 H, d, J═8.7Hz), 7.63 (1 H, d,J═8.7 Hz) and 7.52 (1 H, d, J═8.7 Hz); MS (ESI) m/z:267.0 [M+H]⁺.

[0181] 5,6-Difluoro-N-(6-fluoro-pyridin-3-yl)-nicotinamide (15)

[0182] mp 135-137° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.58 (1 H, brs), 8.52(1 H, s), 8.29-8.23 (1 H, m), 8.03-7.97 (1 H, m), 7.87-7.82 (1 H, m),7.65-7.56 (1 H, m) and 7.19 (1 H,dd, J═8.9, 3.1 Hz); ¹⁹F NMR (282 MHz,DMSO-d₆) δ-73.6 (1 F, d, J═6.5Hz), -132.9 (1 H, q, J═10.7Hz) and -137.1(1 H, q, J═10.7Hz); MS (ESI) m/z: 253.0 [M+H]⁺.

[0183] N-(6-Chloro-pyridin-3-yl)-3-methyl-4-nitro-benzamide (16)

[0184] mp 192-193° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.81 (1 H, brs), 8.74(1 H, d, J═2.6 Hz), 8.21 (1 H, dd, J═8.7, 2.8 Hz), 8.09 (1 H, d, J═8.4Hz), 8.02 (1 H, s), 7.94 (1 H, dd, J═8.5, 1.6 Hz), 7.52 (1 H, d,J═8.7Hz) and 2.58 (3H, s); MS (ESI) m/z: 292.2 [M+H]⁺.

[0185] 5-Fluoro-1 H-indole-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (17)

[0186] mp 290° C.;¹H NMR (300 MHz, DMSO-d₆) δ11.92 (1 H, brs), 10.58 (1H,brs), 8.79 (1 H, s), 8.25 (1 H, dd, J═8.7 and 2.4 Hz), 7.53-7.41 (4 H,m) and 7.10 (1 H, t, J═7.1 Hz); ¹⁹F NMR (282 MHz, DMSO-d₆) δ-123.0 (m);MS (ESI) m/z: 290.0 [M+H]⁺.

[0187] 5-Chloro-1 H-indole-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (18)

[0188] mp 296° C.;¹H NMR (300 MHz, DMSO-d₆) δ12.03 (1 H, brs), 10.62 (1H, brs), 8.80 (1 H, d, J═2.6 Hz), 8.24 (1 H, dd, J═8.7 and 2.6 Hz), 7.79(1 H, s) 7.52 (1 H, d,J═8.7 Hz), 7.42 (1 H, d, J═8.9 Hz), 7.41 (1 H, s)and 7.23 (1 H, dd, J═8.7,1.9 Hz); MS (ESI) m/z: 306.0 [M+H]⁺.

[0189] 5-Chloro-benzofuran-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (19)

[0190] mp 222-223° C.; ¹H NMR (300 MHz, DMSO-d₆) δ11.00 (1 H, brs), 8.78(1 H, d, J ═2.6 Hz), 8.24 (1 H, dd, J═8.7, 2.8 Hz), 7.90 (1 H, d, J═2.1Hz), 7.76 (1 H, s,), 7.73 (1 H, d, J═8.9 Hz) and 7.51 (2 H, d, J═8.9Hz); MS (ESI) m/z: 306.9 [M+H]⁺.

[0191] 5-Chloro-thiophene-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (20)

[0192] mp 215-216° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.61 (1 H, brs), 8.68(1 H, d, J ═2.8 Hz), 8.14 (1 H, dd, J═8.7, 2.6 Hz), 7.88 (1 H, d, J═4.2Hz), 7.50 (1 H, d, J═8.5 Hz) and 7.27 (1 H, d, J═4.0 Hz); ¹³C NMR (75MHz, DMSO-d₆) δ159.7, 144.8, 142.0, 138.5, 135.3, 135.2, 131.5, 130.4,129.0 and 124.7.0; MS (ESI) m/z: 273.0 [M+H]⁺.

[0193] 5-Chloro-furan-2-carboxylic acid (6-chloro-pyridin-3-yl)-amide(21)

[0194] mp 143-144° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.59 (1 H, brs), 8.72(1 H, d, J ═2.4 Hz), 8.18 (1 H, dd, J═8.7, 2.6 Hz), 7.50 (1 H, d, J═8.7Hz), 7.43 (1 H, d,J═3.5 Hz) and 6.76 (1 H, d, J═3.7 Hz); MS (ESI) m/z:257.1 [M+H]⁺. 4.5-Dichloro-isothiazole-3-carboxylic acid(6-chloro-pyridin-3-yl)-amide (22)

[0195] mp 199-201° C.; ¹H NMR (300 MHz, DMSO-d₆) δ11.16 (1 H, brs), 8.75(1 H, d, J ═2.6 Hz), 8.20 (1 H, dd, J═8.7, 2.8 Hz) and 7.51 (1 H, d,J═8.7 Hz); MS (ESI) m/z: 307.9[M+H]⁺.

[0196] 3-Methvl-1 H-indole-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (23)

[0197] mp 184-185° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.64 (1 H, brs), 8.75(1 H, d, J ═2.6 Hz), 8.21 (1 H, dd, J═8.7, 2.8 Hz), 7.68 (1 H, d, J═8.0Hz), 7.54 (1 H, d, J═8.5 Hz), 7.50 (2 H, d, J═8.9 Hz), 7.34 (1 H, s),7.31 (1 H, t, J═7.1 Hz), 7.13 (1 H, t, J═7.1 Hz) and 3.98 (3 H, s); MS(ESI) m/z: 286.1 [M+H]⁺.

[0198] 5-Ethyl-1 H-indole-2-carboxylic acid(6-chloro-pyridin-3-yl)-amide (24)

[0199] mp 270° C.;¹H NMR (300 MHz, DMSO-d₆) δ11.67 (1 H, brs), 10.49 (1H, brs), 8.80 (1 H, s), 8.25 (1 H, dd, J═8.7 and 2.8 Hz), 7.51 (1 H, d,J═8.7 Hz), 7.46 (1 H, s), 7.36 (1 H, d, J═8.7 Hz), 7.35 (1 H, s), 7.10(1 H, d, J═8.5 Hz), 2.66 (2 H, q, J═7.7 Hz) and 1.21 (1 H, t, J═7.7 Hz);¹³C NMR (75 MHz, DMSO-d₆) δ160.6, 144.3, 141.7, 136.2, 136.0, 131.1,127.6, 125.6, 124.7, 120.4, 112.8, 104.8, 28.8 and 16.8; MS (ESI) m/z:300.2 [M+H]⁺.

[0200] N-(6-Chloro-pyridin-3-yl)-benzamide (25)

[0201] mp 163-164° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.59 (1 H, brs), 8.74(1 H, d, J ═2.6 Hz), 8.21 (1 H, dd, J═8.7, 2.8 Hz), 7.94 (1 H, s), 7.92(1 H, d, J═1.6 Hz), 7.59 (1 H, d, J═7.1 Hz) and 7.55-7.49 (3H, m); MS(ESI) m/z: 233.0 [M+H]⁺.

[0202] 1H-Indole-2-carboxylic acid (6-chloro-pyridin-3-yl)-amide (26)

[0203] mp 260-263° C.;¹H NMR (300 MHz, DMSO-d₆) δ11.85 (1 H, brs), 10.62(1 H, brs), 8.78 (1 H, d, J ═2.6 Hz), 8.23 (1 H, dd, J═8.7, 2.8 Hz),7.66 (1 H, d, J═8.0 Hz), 7.51 (1 H, d, J═8.5 Hz), 7.45 (1 H, d, J═8.4Hz), 7.41-7.40 (1 H, m), 7.23 (1 H, t, J═7.1 Hz) and 7.06 (1 H, d, J═7.3Hz); MS (ESI) m/z: 272.0 [M+H]⁺.

[0204] Benzo[b]thiophene-2-carboxvlic acid (6-chloro-pvridin-3-vl)-amide(27)

[0205] mp 226-227° C.; ¹H NMR (300 MHz, DMSO-d6) δ10.86 (1 H, brs), 8.75(1 H, d, J═2.6 Hz), 8.34 (1 H, s), 8.21 (1 H, dd, J═8.7, 2.8 Hz),8.06-7.99 (2 H, m) and 7.54-7.48 (3 H, m); MS (ESI) m/z: 289.1 [M+H]⁺.

[0206] 5-Fluoro-1 H-indole-2-carboxylic acid(6-methyl-prydin-3-yl)-amide (28)

[0207] mp 303-304° C.; ¹H NMR (300 MHz, DMSO-d₆) δ11.88 (1 H, brs),10.36 (1 H, brs), 9.05 (1 H, d, J ═1.9 Hz), 8.06 (1 H, dt, J═8.4, 2.1Hz), 7.45 (2 H, m), 7.39 (1 H, s), 7.24 (1 H, d, J═8.3 Hz), 7.08 (1 H,dt, J═9.2, 2.4 Hz) and 2.43 (3 H, s); ¹⁹F NMR (282 MHz, DMSO-d₆)δ-123.2; MS (ESI) m/z: 270.2 [M+H]⁺.

[0208] 3,4-Difluoro-N-(6-trifluoromethyl-pvridin-3-vl)-benzamide (29)

[0209] mp 175-176° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.84 (1 H, brs), 9.05(1 H, d, J ═1.8 Hz), 8.45 (1 H, dd, J═8.5, 1.9 Hz), 8.07 (1 H, ddd,J═9.9, 7.6, 2.1 Hz), 7.93 (1 H, d, J═8.5 Hz) and 7.89-7.86 (1 H, m); 1⁹FNMR (282 MHz, DMSO-d₆) 6 -65.7 (3F, s), -132.5 (IF, m) and -137.0 (1 F,m); MS (ESI) m/z: 303.1 [M+H]⁺.

[0210] N-(6-Chloro-pvridin-3-yl)-3-fluoro-4-1-pyrrolidin-1-yl-benzamide(30)

[0211] mp 215-216° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.23 (1 H, brs), 8.71(1 H, d, J ═2.6 Hz), 8.18 (1 H, dd, J═8.7, 2.4Hz), 7.70 (1 H, s), 7.65(1 H, d, J═3.1 Hz), 7.46 (1 H, d, J═8.9 Hz), 6.73 (1 H, t, J═8.5 Hz),3.42-3.40 (4 H, m) and 1.91-1.88 (4H, m); ¹⁹F NMR (282 MHz, DMSO-d₆)δ-129.0 (s); MS (ESI) m/z: 320.2 [M+H]⁺.

[0212] N-(6-Chloro-pvridin-3-yl)-3-fluoro-4-morpholin-4-yl-benzamide(31)

[0213] mp 228-229° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.41 (1 H, s), 8.75(1 H, d, J═2.6 Hz), 8.20 (1 H, dd, J═8.7 and 2.6 Hz), 7.79 (1 H, s),7.74 (1 H, d, J═7.7 Hz), 7.50 (1 H, d, J═8.7 Hz), 7.13 (1 H, t, J═9.2Hz), 3.74 (4 H, m) and 3.12 (4 H, m); ¹³C NMR (75 MHz, DMSO-d₆) δ164.8,155.7, 152.5, 144.4, 143.3 (d, J═7 Hz), 142.0, 135.9, 131.4, 127.3 (d,J═7Hz), 125.5, 124.6, 118.7 (d, J═3 Hz), 66.5 and 50.4 (d, J═5 Hz); MS(ESI) m/z: 336.2 [M+H]⁺.

[0214] N-(6-Chloro-pyrdin-3-yl)-3-fluoro-4-imidazol-1-yl-benzamide (32):

[0215] mp 215-218° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.71 (1 H ,s), 8.78 (1H, s), 8.24 (1 H, dt, J═8.7, 1.6 Hz), 8.17 (1 H, s), 8.07 (1 H, dd,J═12.0, 1.6 Hz), 7.96 (1 H, dd, J═8.4, 1.7 Hz), 7.88 (1 H, t, J═8.2 Hz),7.69 (1 H, s), 7.53 (1 H, d, J═8.7 Hz) and 7.17 (1 H, s); ¹⁹F NMR (282MHz, DMSO-d₆) δ-123.1 (t, J═8.8 Hz); MS (ESI) m/z: 317.1 [M+H]⁺.

[0216]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-[(pyridin-2-ylmethyl)-amino]-benzamide(33)

[0217] mp 210-211° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.20 (1 H, brs), 8.69(1 H, d, J ═2.6 Hz), 8.50 (1 H, d, J═4.7 Hz), 8.15 (1 H, dd, J═8.7, 2.8Hz), 7.75-7.64 (2 H, m), 7.45 (1 H, d, J═8.7 Hz), 7.31 (1 H, d, J═8.0Hz), 7.25 (1 H, dd, J═6.4, 5.1Hz), 6.63 (1 H, t, J═8.7 Hz) and 4.49 (2H, d, J═5.9 Hz); ¹⁹F NMR (282 MHz, DMSO-d₆) δ-134.3 (m); MS (ESI) m/z:357.0 [M+H]⁺.

[0218] N-(6-Chloro-pyridin-3-yl)-4-dimethylamino-3-fluoro-benzamide (34)

[0219] mp 170-171° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.32 (1 H, brs), 8.76(1 H, s), 8.21 (1 H, d, J═8.7 Hz), 7.75 (1 H, s), 7.71 (1 H, d, J═5.8Hz), 7.59 (1 H, d, J═8.7 Hz), 7.02 (1 H, t, J═9.2 Hz) and 2.92 (6 H, s);¹⁹F NMR (282 MHz, DMSO-d₆) δ-122.6 (t, J═10.7 Hz); MS (ESI) m/z: 294.2[M+H]⁺.

[0220] Pyridine-2-carboxylic acid[4-(6-chloro-pyridin-3-ylcarbamoyl)-phenyl]-amide (35)

[0221] mp 258-260° C.;¹H NMR (300 MHz, DMSO-d6) δ10.92 (1 H, s), 10.50(1 H, s), 8.78 (1 H, d, J═2.6 Hz),), 8.75 (1 H, d, J═4.7 Hz), 8.24 (1 H,dd, J═8.7 and 2.6 Hz), 8.17 (1 H, d, J═7.8 Hz), 8.09 (2 H, d, J═7.8 Hz),8.08 (1 H, m), 7.98 (2 H, d, J═8.7 Hz), 7.70 (1 H, dd, J═6.4, 5.0 Hz)and 7.50 (1 H, d, J═8.7 Hz); ¹³C NMR (75 MHz, DMSO-d₆) δ165.8, 163.5,150.0, 149.0, 144.4, 142.3, 141.9, 138.8, 136.1, 131.4, 129.4, 129.2,127.7, 124.6, 123.1 and 120.1; MS (ESI) m/z: 353.2 [M+H]⁺.

[0222] N-(6-Chloro-pvrndin-3-yl)-4-(morpholine-4-sulfonyl)-benzamide(36)

[0223]¹H NMR (300 MHz, DMSO-d₆) δ10.88 (1 H, s), 8.78 (1 H, d, J═2.6Hz), 8.24 (1 H, dd, J═8.7 and 2.6 Hz), 8.19 (2 H, d, J═8.5 Hz), 7.90 (2H, d, J═8.5 Hz), 7.50 (1 H, d, J═8.7 Hz), 3.62 (4H, m) and 2.90 (4H, m);¹³C NMR (75 MHz, DMSO-d₆) δ165.3, 144.9, 142.1, 138.8, 137.9, 135.7,131.5, 129.4, 128.3, 124.7, 65.8 and 46.8; MS (ESI) m/z: 382.1 [M+H]⁺.

[0224] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-nitro-benzamide (37)

[0225] mp 205° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.87 (1 H, brs), 8.69 (1H, d, J ═2.4 Hz), 8.32 (1 H, t, J═7.8 Hz), 8.22 (1 H, dd, J═8.7,2.6Hz),8.10 (1 H, dd, J═11.8, 1.6Hz), 7.96 (1 H, d, J═8.5Hz) and 7.53 (1 H, d,J═8.7 Hz); MS (ESI) m/z: 294.0 [M-H]⁺.

[0226] N-(6-Chloro-pyridin-3-yl)-4-hvdroxyamino-benzamide (38)

[0227] dec 200° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.24 (1 H, brs), 8.86 (1H, s), 8.75 (1 H, d, J ═2.4 Hz), 8.61 (1 H, s), 8.22 (1 H, dd, J═8.7,2.8 Hz), 7.83 (2 H, d, J═8.5 Hz) and 7.47 (1 H, d, J═8.7 Hz); MS (ESI)m/z: 261.9 [M-H]⁺.

[0228] N-(6-Chloro-pvridin-3-yl)-3-fluoro-4-hydroxyamino-benzamide (39)

[0229]¹H NMR (300 MHz, DMSO-d₆) δ10.16 (1 H, brs), 8.74 (1 H, d, J═2.6Hz), 8.19 (1 H, dt, J═8.7, 2.8 Hz), 7.66 (1 H, dd, J═12.7,1.7 Hz), 7.60(1 H, d, J═8.4 Hz), 7.47 (1 H, d, J═8.7 Hz), 6.80 (1 H, t, J═8.9 Hz) and5.93 (2 H, brs); MS (ESI) m/z: 280.0 [M-H]⁺.

[0230] N-(6-Chloro-pvridin-3-vl)-4-methvlsulfamovl-benzamide (40)

[0231] mp 186-189° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.78 (1 H, brs), 8.77(1 H, d, J═2.3 Hz), 8.23 (1 H, dt, J═8.7, 2.8 Hz), 8.13 (2 H, d, J═8.4Hz), 7.99 (1 H, d, J═8.2 Hz), 7.63 (1 H, q, J═5.2 Hz), 7.53 (1 H, d,J═8.7 Hz) and 2.43 (3H, s); MS (ESI) m/z: 326.2 [M+H]⁺.

[0232] 4-Amino-N-(6-chloro-pyridin-3-vl)-3-fluoro-benzamide (41)

[0233] mp 193° C.; ¹H NMR (300 MHz, DMSO-d₆) δ10.16 (1 H, brs), 8.73 (1H, d, J ═2.6 Hz), 8.20 (1 H, dd, J═8.7, 2.8 Hz), 7.66 (1 H, dd,J═12.7,1.7Hz), 7.59 (1 H, dd, J═8.4,1.9Hz), 7.46 (1 H, d, J═8.7 Hz),6.80 (1 H, t, J═8.7 Hz) and 5.94 (2 H, s); MS (ESI) m/z: 266.0 [M+H]⁺.

[0234] N-(6-Chloro-pyridin-3-vl)-4-nitro-benzamide (42)

[0235] mp 193° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.88 (1 H, brs), 8.77 (1H, d, J═2.3 Hz), 8.39 (1 H, s), 8.36 (1 H, s), 8.23 (1 H, dt, J═8.7, 2.8Hz), 8.20 (1 H, s), 8.17 (1 H, s), 7.63 (1 H, q, J═5.2 Hz) and 7.54 (1H, d, J═8.7 Hz): MS (ESI) m/z: 275.9 [M+H]⁺.

[0236] N-(6-Chloro-pyddin-3-yl)-3-chloro-4-fluoro-benzamide (43)

[0237] mp 173° C.;¹H NMR (300 MHz, DMSO-d₆) δ10.65 (1 H, brs), 8.74 (1H, s), 8.23-8.18 (2 H, m), 8.01-7.97 (1 H, m), 7.61 (1 H, t, J═9.1 Hz)and 7.52 (1 H, d, J═8.7 Hz): MS (ESI) m/z: 285.0 [M+H]⁺.

EXAMPLE 9

[0238] Example 9 sets forth representative methods for elaborating thering structures of the substituted benzanilides of the invention

[0239] 9.1 Synthesis of Amino-Modified Pyridyl-Benzanilides

[0240] 9.1a General Method for XX When X ═Cl

[0241] An aniline XX (1.55 mmol) was dissolved in dry THF (5 mL) andtreated with pyridine (0.075 mL, 0.6 equivalents) followed by a4-fluorobenzoyl chloride XIX (1. 1 equivalents; slow addition; aprecipitate forms immediately). After 2 h, saturated NaHCO₃ solution (15mL) was added. After 30 min of vigorous stirring, the mixture wasdiluted with EtOAc (15 mL). The organic layer was washed with water (lx5nlL), dried (Na₂SO₄) and filtered. Removal of the solvent provided theproducts XXI as solids, in high yields (>95%) and with high purity(typically >95% by LC/MS).

[0242] 9.1b General Method for XX When X═OH

[0243] A 4-fluorobenzoic acid XIX (1.7 mmol, 1.1 equivalents) wasdwassolved in dry THF (5 mL) and treated with dry DMF (3 drops) followedby oxalyl chloride (1.3 equivalents; slow addition; gas evolution; mildexotherm). After 1 h, an aniline XX (1.55 mmol) was added (a precipitateforms immediately) and 2 h later, saturated NaHCO₃ solution (15 mL) wasadded. After 30 min of vigorous stirring, the mixture was diluted withEtOAc (15 mL). The organic layer was washed with water (1×5 niL) andsilica gel (2 g) was added. Removal of the solvent provided a solid,which was applied to a column of silica gel (1″×641 ) and eluted withEtOAc/hexanes. The products XXI were thus obtained as solids in goodyields (>70%) and with high purity (>95% by LC/MS).

[0244] 9.1c General Method for Preparing Compounds of Motif XXII

[0245] 5 Compound XXI (0.10 mmol) and RRNH (3 equivalents) were heatedat 110-120° C. in dry DMSO (0.3 mL) for 20 h. The mixture was brought toroom temperature and diluted with water (0.3 mL), saturated NaHCO₃solution (0.3 mL), EtOAc (4 mL) and brine (2 mL). The organic layer waswashed with brine (2×1 mL) and silica gel (400 mg) was added. Removal ofthe solvent provided a solid, which was applied to a column of silicagel (0.5−×6″) and eluted with EtOAc/hexanes. The products XXII wereobtained as solids in good yields (>65%) and with high purity (>95% byLC/MS).

[0246] 9.2 Synthesis of Aryl-Modified Benzanilides

[0247] 9.2a General Method for Preparing XXIII

[0248] Compound XXI (0.15 mmol), a phenol (1.2 equivalents) and K₂CO₃were heated at 110-120° C. in dry DMSO (0.5 mL) for 20 h. The mixturewas brought to room temperature and diluted with water (0.7 mL), EtOAc(3 mL) and brine (1 mL). The organic layer was washed with brine (2×0.5mL) & silica gel (400 mg) was added. Removal of the solvent provided asolid, which was applied to a column of silica gel (0.5″×6″) and elutedwith EtOAc/hexanes. The products XXIII were obtained as solids in goodyields (>70%) and with high purity (>95% by LC/MS).

[0249] 9.3 Preparation of Thioether-Modified Benzanilides

[0250] 9.3a General Method for Preparing Compounds of Motif XXIV

[0251] A 0.2-0.4 M suspension of XXI in DMSO, Cs₂CO₃ (1-2 eq.) and alkylor arylsulfide (1.2-3 eq.) was agitated at rt or up to 75° C. for 4 h to2 days until the reaction was judged to have gone to completion (TLC orLCMS). The reaction mixture was partitioned between ethyl acetate andwater. The aqueous layer was washed with water, 1 N NaOH solution (2x),water, and brine, dried (Na₂SO₄) and concentrated. Chromatography onSiO₂, trituration or recrystallization afforded products XXIV as solidsin good yields (>70%) and with high purities (>95% by LC/MS).

[0252] 9.4 Preparation of Oxidized Thioether-Modified Benzanilides

[0253] 9.4a General Method forPreparing Compounds of Motif XXV

[0254] A 0. 1-0.25 M solution of sulfide XXIV in 2:1 CH₂CI₂/THF wastreated with mCPBA (2 eq.) and agitated at rt overnight. The reactionmixture was concentrated in vacuo and the residue was partitionedbetween ethyl acetate and water. The organic layer was washed with 1 NNaOH solution (3x), water, and brine, dried (Na₂SO₄) and concentrated.Chromatography on SiO₂, trituration or recrystallization afforded pureproduct XXV as solids in good yields (>70%) and with high purity (>95%by LC/MS).

EXAMPLE 10

[0255] Example 10 sets forth the results of the characterization ofrepresentative compounds prepared by the methods of Examples 8 and 9.The compounds were characterized using a combination of melting point,¹H NMR and mass spectrometry. The results of the characterization arepresented below. The structures for the compounds set forth below areprovided in FIG. 1.

[0256] N-(6-Chloropyridin-3-yl)-3-fluoro-4-(2-phenethylamino)-benzamide(156)

[0257]¹H NMR (CDCl₃, CD₃OD) δ8.40 (d, J═2.4 Hz, 1 H), 8.32 (dd, J═2.6,8.7 Hz, 1 H), 7.54-7.61 (m, 2 H), 7.19-7.34 (m, 6 H), 6.70 (t, J═8.0 Hz,1 H), 3.47 (t, J ═7.0 Hz, 2 H), 2.94 (t, J═7.0 Hz, 2 H).

[0258]N-(6-Chloropyridin-3-yl)-3-fluoro-4-(2-(2-pyridyl)ethylamino)-benzamide(122) ¹H NMR (CDCl₃, CD₃OD) δ8.51 (dd, J═1.6, 5.7 Hz, 1 H), 8.41 (d, J═2.4 Hz, 1 H), 8.31 (dd, J═2.8, 8.7 Hz, 1 H), 7.53-7.66 (m, 3H), 7.28(d, J═9.0 Hz, 1 H), 7.15-7.19 (m, 2 H), 6.72 (t, J═8.5 Hz, 1 H), 3.60(t, J ═7.0 Hz, 2 H), 3.10 (t, J═6.9 Hz, 2 H).

[0259] N-(6-Chloropyridin-3-yl)-3-fluoro-4-(2-(3-pyridl)ethylamino)-benzamide (123)

[0260]¹H NMR (CDCl₃, CD₃OD) δ8.39-8.41 (m, 3 H), 8.30 (dd, J═2.6, 8.7Hz, 1 H), 7.53-7.63 (m, 3 H), 7.23-7.28 (m, 2 H), 6.66 (t, J═8.2 Hz, 1H), 3.48 (t, J ═7.0 Hz, 2 H), 2.93 (t, J═7.0 Hz, 2 H).

[0261]N-(6-Chloropyridin-3-yl)-3-fluoro-4-(2-(4-pyridyl)ethylamino)-benzamide(124)

[0262]¹H NMR (CDCl₃, CD₃OD) Δ8.43 (d, J═2.1 Hz, 2 H), 8.39 (d, J ═2.5Hz, 1 H), 8.32 (dd, J═2.8, 8.7 Hz, 1 H), 7.56-7.63 (m, 2 H), 7.27 (d, J═9 Hz, 1 H), 7.15 (d, J ═6.0 Hz, 1 H), 6.66 (t, J═8.4 Hz, 1 H), 3.50 (t,J═7.2 Hz, 2 H), 2.93 (t, J ═7.0 Hz, 2 H).

[0263]N-(6-Chloropyridin-3-yl)-4-(3,4-difluorophenoxy)-3-fluorobenzamide (210)

[0264]¹H NMR (CDCl₃, CD₃OD) o 8.42 (d, J═2.6 Hz, 1 H), 8.33 (dd, J═2.6,4.5 Hz, 1 H), 7.80 (dd, J═2.1, 11.2 Hz, 1 H), 7.70 (dd, J═1.0, 8.5 Hz, 1H), 7.30 (d, J ═8.7 Hz, 1 H), 7.00-7.17 (m, 2 H), 6.72-6.88 (m, 2 H);MS(ESI): 379 (MH⁺).

[0265]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(furan-2-ylmethylsulfanl)-benzamide(134)

[0266]¹H NMR (DMSO-d₆) δ10.56 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═8.7Hz, 1 H), 7.82-7.74 (m, 2 H), 7.67 (dd, J ═7.8, 8.0, 1H), 7.57 (s, 1 H),7.51 (d, J═8.5 Hz, 1 H), 6.38-6.32 (m, 2 H), 4.42 (s, 2 H).

[0267] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-phenethylsulfanyl-benzamide(135) ¹H NMR (DMSO-d₆) δ10.55 (s, 1 H), 8.77 (s, 1 H), 8.22 (d, J═8.5Hz, 1 H), 7.85-7.75 (m, 2 H), 7.61 (dd, J═7.9, 7.9 Hz, 1 H), 7.51 (d,J═8.7 Hz, 1 H), 7.36-7.18 (m, 5 H), 3.39-3.22 (m, 2 H), 2.92 (t, J═7.7Hz, 2 H).

[0268]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-phenyl-ethanesulfonyl)-benzamide(141)

[0269]¹H NMR (DMSO-d6) δ10.82 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═8.7Hz, 1 H), 8.05 (d, J═10.7 Hz, 1 H), 7.93 (d, J═8.2 Hz, IH), 7.81 (dd,J═7.1, 7.1 Hz, IH), 7.62-7.52 (m, 3H), 7.19 (d, J═8.2 Hz, 1 H), 4.92 (s,2 H)

[0270]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-phenylmethanesulfonyl-benzamide(142)

[0271]¹H NMR (DMSO-d₆) δ10.56 (s, 1 H), 8.75 (s, 1 H), 8.19 (d, J═8.5,1H), 7.76 (d, J═9.4,2H), 7.66-7.34 (m, 4 H), 7.18-7.06 (m, 2 H), 4.36(s, 2 H)

[0272]N-(6-Chloro-pyridin-3-yl1-3-fluoro-4-(thiophen-2-ylmethanesulfonyl)-benzamide(146)

[0273]¹H NMR (DMSO-d₆) δ10.57 (s, 1 H), 8.76 (s, 1 H), 8.21 (d, J═8.5Hz, IH), 7.84-7.58 (m, 4 H), 7.52 (d, J═8.3 Hz, 1 H), 7.40 (d, J═5.4 Hz,IH), 7.07 (s, 1 H), 6.93 (d, J═4.9 Hz, 1 H), 4.64 (s, 2 H).

[0274]N-(6-Chloro-pyridin-3-yl1-3-fluoro-4-(2-py)din-2-yl-ethanesulfonyl)-benzamide(147)

[0275]¹H NMR (DMSO-d6) δ10.56 (s, 1 H), 8.76 (d, J═2.3 Hz, 1 H), 8.51(d, J═4.5 Hz, IH), 8.22 (dd, J═2.4, 8.7 Hz, 1 H), 7.85-7.68 (m, 3H),7.63 (dd, J═7.6, 8.2 Hz, 1 H), 7.51 (d, J═8.7 Hz, IH), 7.33 (d, J═7.8Hz, 1 H), 7.25 (dd, J═5.6, 6.6, 1H), 3.47 (t, J═7.3 Hz, 1 H), 3.09 (t,J═7.5 Hz, 2 H).

[0276]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-pyridin-4-yl-ethanesulfonyl)-benzamide(155)

[0277]¹H NMR (DMSO-d₆) δ10.55 (s, 1 H), 8.76 (s, 1 H), 8.47 (d, J═5.2,2H), 8.22 (d, J═8.7 Hz, 1 H), 7.87-7.74 (m, 2 H), 7.62 (dd, J═7.8, 8.2Hz, 1 H), 7.49 (d, J═8.7, 1H), 7.30 (d, J═5.3 Hz, 2 H), 3.40 (t, J═7.3Hz, 2 H), 2.94 (t, J═7.4 Hz, 2 H).

[0278]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(thiophen-2-vlmethanesulfonyl)-benzamide(150)

[0279]¹H NMR (DMSO-d₆) δ10.84 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═6.1Hz, 1 H), 8.04 (d, J═10.6 Hz, 1 H), 7.91 (d, J═8.0 Hz, 1 H), 7.83 (d,J═7.7 Hz, 1 H), 7.58-7.46 (m, 2 H), 7.0-6.95 (m, 2 H), 5.12 (s, 2 H).

[0280]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(4-fluoro-phenylmethanesulfonyl)-benzamide(151)

[0281]¹H NMR (DMSO-d₆) δ10.82 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═8.7Hz, 1 H), 7.90 (d, J═8.2 Hz, 1 H), 7.76 (dd, J═7.1, 7.9 Hz, 1 H), 7.55(d, J═8.7 Hz, 1 H), 7.30-7.23 (m, 2 H), 7.15 (dd, J═8.7, 8.9 Hz, 2 H),4.85 (s, 2 H)

[0282]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(pyridin-2-ylsulfanyl)-benzamide(157)

[0283]¹H NMR (DMSO-d₆) δ10.71 (s, 1 H), 8.78 (d, J═2.3 Hz, 1 H), 8.37(d, J═4.0, 1H), 8.23 (dd, J═2.5, 8.6 Hz, 1 H), 7.95-7.67 (m, 4 H), 7.53(d, J═8.7, 1H), 7.28-7.16 (m, 2 H).

[0284]4-(2-Chloro-phenlsulfanyl)-N-(6-chloro-pyridin-3-yl)-3-fluoro-benzamide(158)

[0285]¹H NMR (DMSO-d₆) δ10.65 (s, 1 H), 8.76 (d, J═2.6 Hz, 1 H), 8.21(dd, J═2.6, 8.7 Hz, 1 H), 7.91 (d, J═10.4 Hz, 1 H), 7.80 (d, J═8.1 Hz, 1H), 7.62 (d, J═7.4 Hz, 1 H), 7.52 (d, J═8.7 Hz, 1 H), 7.44-7.24 (m, 4H).

[0286]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-fluoro-phenylsulfanyl)-benzamide(159)

[0287]¹H NMR (DMSO-d₆) δ10.61 (s, 1 H), 8.74 (d, J═2.4 Hz, 1 H), 8.20(dd, J═2.6, 8.7, 1 H), 7.87 (d, J═10.6, 1 H), 7.72 (d, J═10.2 Hz, 1 H),7.60-7.45 (m, 3H), 7.40 (dd, J═8.7, 9.0 Hz, 1 H), 7.30 (dd, J═7.6, 7.7Hz, 1 H), 7.18 (dd, J═7.8, 7.9 Hz, 1 H)

[0288] 4-tert-Butylsulfanyl-N-(6-chloro-pyridin-3-yl)-3-fluoro-benzamide(160)

[0289]¹H NMR (DMSO-d₆) δ10.69 (s, 1 H), 8.78 (s, 1 H), 8.23 (d, J═6.1Hz, 1 H), 7.92-7.70 (m, 3 H), 5.53 (d, J═8.4 Hz, 1 H), 1.29 (s, 9 H).

[0290] 4-Butylsulfanyl-N-(6-chloro-pyridin-3-yl)-3-fluoro-benzamide(161)

[0291]¹H NMR (DMSO-d₆) δ10.56 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═6.3Hz, 1 H), 7.88-7.70 (m, 2 H), 7.60-7.46 (m, 2 H), 3.07 (t, J═7.2 Hz, 2H), 1.63-1.50 (m, 2 H), 1.48-1.28 (m, 2 H), 0.89 (t, J═7.0 Hz, 3 H).

[0292] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-isobutylsulfanyl-benzamide(163)

[0293]¹H NMR (DMSO-d₆) δ10.55 (s, 1 H), 8.77 (s, 1 H), 8.22 (d, J═6.9Hz, 1 H), 7.83-7.72 (m, 2 H), 7.60-7.44 (m, 2 H), 2.96 (d, J═6.5 Hz, 2H), 1.88-1.70 (m, 1 H), 1.01 (d, J═6.4 Hz, 6 H).

[0294]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(pyridin-4-ylsulfanyl)-benzamide(164)

[0295]¹H NMR (DMSO-d₆) δ10.73 (s, 1 H), 8.78 (s, 1 H), 8.42 (d, J═4.2Hz, 2 H), 8.23 (d, J═8.7 Hz, 1 H), 7.98 (d, J═9.9 Hz, 1 H), 7.91 (d,J═8.2 Hz), 7.82 (dd, J═7.3, 7.8 Hz, 1 H), 7.53 (d, J═8.7Hz, 1 H), 7.12(d, J═5.7Hz, 2 H)

[0296]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(4-fluoro-phenylsulfanl)-benzamide(165)

[0297]¹H NMR [(CD₃) ₂CO)]δ9.98 (s, 1 H), 8.75 (d, J═2.4 Hz, 1 H), 8.37(dd, J═1.2, 2.8 Hz, 1 H), 7.84-7.74 (m, 2 H), 7.66-7.57 (m, 2 H), 7.50(d, J═8.7 Hz, 1 H), 7.34-7.25 (m, 2 H), 7.14 (dd, J═7.6, 8.0 Hz, 1 H).

[0298]4-(2-Chloro-benzenesulfonyl)-N-(6-chloro-pyridin-3-yl)-3-fluoro-benzamide(166)

[0299]¹H NMR [(CD₃)₂CO)]δ10.09 (s, 1 H), 8.76 (d, J═2.7 Hz, 1 H),8.43-8.29 (m, 3 H), 8.11 (dd, J═1.6, 8.2 Hz, 1 H), 7.87 (dd, J═1.5, 10.7Hz, 1 H), 7.84-7.71 (m, 2 H), 7.64 (dd, J═1.4, 7.6 Hz, 1 H), 7.47 (d,J═8.7 Hz, 1 H).

[0300] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-fluoro-benzenesulfonyl)-benzamide (167)

[0301]¹H NMR [(CD₃) ₂CO)]δ10.09 (s, 1 H), 8.76 (d, J═2.7 Hz, 1 H),8.35-8.26 (m, 2 H), 8.19 (dd, J═7.3, 7.7 Hz, 1 H), 8.11 (d, J═8.0 Hz, 1H), 7.91-7.82 (m, 2 H), 7.58 (dd, J═7.6, 7.7 Hz, 1 H), 7.48 (d, J═8.7Hz, 1 H), 7.37 (dd, J═8.5, 10.3 Hz, 1 H).

[0302]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-methyl-propane-2-sulfonyl)-benzamide(168)

[0303]¹H NMR (DMSO-d₆) δ8.77 (s, 1 H), 8.23 (d, J═8.7 Hz, 1 H),8.08-7.85 (m, 3H), 7.55 (d, J═8.9 Hz, 1 H), 1.30 (s, 9 H).

[0304]4-(Butane-1-sulfonyl)-N-(6-chloro-pyridin-3-yl)-3-fluoro-benzamide (169)

[0305]¹H NMR (DMSO-d₆) δ10.85 (s, 1 H), 8.77 (s, 1 H), 8.21 (s, 1 H),8.08-7.86 (m, 3H), 7.55 (d, J═9.1 Hz, 1 H), 3.43 (t, J═7.5 Hz, 2 H),1.60-1.44 (m, 2 H), 1.41-1.23 (m, 2 H), 0.84 (t, J═7.0 Hz, 3 H).

[0306]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(propane-2-sulfonyl)-benzamide(170)

[0307]¹H NMR (DMSO-d₆) δ10.85 (s, 1 H), 8.76 (s, 1 H), 8.22 (d, J═8.5Hz, 1 H), 8.07-7.92 (m, 3 H), 7.55 (d, J═8.7 Hz, 1 H), 3.58-3.48 (m, 1H), 1.22 (d, J═6.8 Hz, 6 H).

[0308]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-methyl-propane-1-sulfonyl)-benzamide(171)

[0309]¹H NMR (DMSO-d₆) δ8.77 (s, 1 H), 8.23 (d, J═8.7 Hz, 1 H),8.06-7.98 (m, 3H), 7.55 (d, J═8.9 Hz, 1 H), 3.38 (d, J═6.2 Hz, 2 H),2.12-2.00 (m, 1 H), 0.99 (d, J═6.8 Hz, 6 H).

[0310] N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(pyrazin-2-ylmethylsulfanyl)-benzamide (175)

[0311]¹H NMR (DMSO-d₆) δ10.55 (s, 1 H), 8.75(d, J═2.4 Hz, 1 H), 8.72 (s,1 H), 8.55 (s, 1 H), 8.51(d, J═2.5 Hz, 1 H), 8.20 (dd, J═2.5, 8.6 Hz),7.82-7.72 (m, 2 H), 7.68 (dd, J═7.7, 7.9 Hz, 1 H), 7.50 (d, J═8.7, 1 H),4.53 (s, 2 H).

[0312]N-(6-Chloro-pyridin-3-yl)-3-fluoro-4-(2-pyrazin-2-yl-ethylsulfanl)-benzamide(176)

[0313]¹H NMR (DMSO-d₆) δ10.56 (s, 1 H), 8.77 (s, 1 H), 8.59 (s, 1 H),8.57 (s, 1 H), 8.49 (s, 1 H), 8.22 (d, J═8.9 Hz, 1 H), 7.88-7.74 (m, 2H), 7.63 (dd, J═8.0, 8.0 Hz, 1 H), 7.51 (d, J═8.7 Hz, 1 H), 3.50 (t,J═7.3, 2H), 3.14 (t, J═7.3 Hz, 2 H).

EXAMPLE 11

[0314] This example illustrates a KCNQ2 screening protocol forevaluating compounds of the present invention.

[0315] Cells expressing voltage-gated K+ channels, such as KCNQ2-likechannels were loaded with ⁸⁶Rb+ by culture in media containing ⁸⁶RbCl.Following loading, culture media was removed and the cells were washedin EBSS to remove residual traces of ⁸⁶Rb+. Cells were preincubated withdrug (0.01 - 30 pM in EBSS) and then ⁸⁶Rb+ efflux was stimulated byexposing cells to EBSS solution supplemented with a sub-maximalconcentration of KCl (generally 7-20 mM) in the continued presence ofdrug. After a suitable efflux period, the EBSS/ KC1 solution was removedfrom the cells and the ⁸⁶Rb+ content determined by Cherenkov counting(Wallac Trilux). Cells were then lysed with a SDS solution and the ⁸⁶Rb+content of the lysate determined. Percent ⁸⁶Rb+ efflux was calculatedaccording to:

(⁸⁶Rb+ content in EBSS/(⁸⁶Rb+ content in EBSS+ ⁸⁶Rb+ content of thelysate))*100

[0316] Efflux was normalized to the maximal ⁸⁶Rb+ efflux (i.e., thatinduced by a high concentration of KCl, generally 30-135 mM).

[0317] The compounds of the invention (FIG.1) were prepared according tothe general methods set forth in the examples and they were assayedusing the above-described method. The activity of the assayed compoundsranged from about 30% to greater than about 70% efflux.

[0318] It is understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication and scope of the appended claims. All publications, patents,and patent applications cited herein are hereby incorporated byreference in their entirety for all purposes.

What is claimed is:
 1. A compound having the formula:

wherein Y is a member selected from H, methyl, methoxy,trifluoromethoxy, —CF₃ or halo; V and X are members independentlyselected from H, halo, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, NO₂, CN, CF₃,C(O)NR¹¹R¹² and C(O)R¹³; R¹, R¹¹, R¹² and R¹³ are members independentlyselected from substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, substituted or unsubstitutedcarbocycle, substituted or unsubstituted heterocycle, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹¹ and R¹² optionally can be joined into a ring; Q and W are membersindependently selected from —(CR²R³)_(t)—(CH₂)_(n)—,—(CH₂)_(n)—(CR²R³)_(t), —C(R⁴)═C(R⁵)—, and —C≡C— wherein R² and R³ aremembers independently selected from H, F, substituted or unsubstitutedlower alkyl or substituted or unsubstituted lower heteroalkyl, in whichR² and R3 are optionally joined to form a cyclic structure which is amember selected from the group consisting of cycloalkyl and heterocyclegroups, or R² and R³ together with the carbon to which they are attachedform —C(O)—; Z represents —O—, —S(O)m—, —N(R⁴)—, —N(⁴)C(O)—,—C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—, —N(R⁴)C(O)O—, (CR²R³)_(t), and—SO₂N(R⁴)—, wherein R⁴ and R⁵ are members independently selected fromthe group consisting of H, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹ is optionally joined together with either X or R⁴ to form asubstituted or unsubstituted heterocycle; m is an integer from 0 to 2,inclusive; n is an integer from 0 to 3, inclusive; and t is an integerfrom 0 to 2, inclusive.
 2. The compound according to claim 1 , wherein Yis a member selected from chloro and methyl.
 3. The compound accordingto claim 1 , wherein V and X are members independently selected from thegroup consisting of H, halo, substituted or unsubstituted lower alkyl,and —CF₃.
 4. The compound according to claim 1 , wherein Z is a memberselected from the group consisting of —S—, SO₂—, —(CR²R³)_(t)—, and —O—.5. The compound according to claim 3 , wherein Z is a member selectedfrom the group consisting of —S—, SO₂—, —(CR²R³)_(t)—, and —O—.
 6. Thecompound according to claim 1 , wherein R⁴ is H.
 7. The compoundaccording to claim 3 , wherein R¹ is a member selected from the groupconsisting of:


8. The compound according to claim 3 , wherein n is an integer from 0 to2, inclusive; and t is an integer from 0 to 1, inclusive.
 9. Thecompound according to claim 1 , having a structure which is a memberselected from the group consisting of the compounds set forth in FIG. 1.10. A compound according to claim 1 , having the formula:

wherein R¹—W—Z—Q— is R⁶, and R⁶ is selected from the group consisting ofH, halogen, substituted or unsubstituted alkyl, halo(C₁-C₄)alkyl, nitro,cyano, substituted or unsubstituted phenyl, R⁹O—; R⁹S—; R⁹NH—; R⁹NH—;R⁹NHS(O)₂—; R⁹S())₂—, with the proviso that both X and R⁶ are not H;wherein R⁹ is a member selected from aryl, and alkylaryl, when there ismore than one R⁹ group per molecule, each R⁹ group is independentlyselected; and Y is a member selected from halogen, C₁-C₄ alkyl, —OCH₃,and —OCF₃.
 11. The compound according to claim 10 , wherein the alkylcomponent of said alkylaryl group is a C₁-C₄ alkyl group.
 12. Thecompound according to claim 10 , wherein said aryl group of R⁹ isheteroaryl.
 13. The compound according to claim 10 , wherein the arylcomponent of said (C₁-C₄)alkylaryl group is a substituted orunsubstituted aryl group.
 14. The compound according to claim 10 ,wherein the aryl component of said (C₁-C₄)alkylaryl group is asubstituted or unsubstituted heteroaryl group.
 15. A method ofincreasing ion flow through voltage-dependent potassium channels in acell, said method comprising contacting said cell with a potassiumchannel-opening amount of a compound of the formula:

wherein Y is a member selected from H, methyl, methoxy,trifluoromethoxy, —CF₃ or halo; V and X are members independentlyselected from H, halo, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, NO₂, CN, CF₃,C(O)NR¹¹R¹² and C(O)R¹³; R¹, R¹¹, R¹² and R¹³ are members independentlyselected from substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, substituted or unsubstitutedcarbocycle, substituted or unsubstituted heterocycle, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹¹ and R¹² optionally can be joined into a ring; Q and W are membersindependently selected from —(CR²R³)_(t)—(CH₂)_(n)—,—(CH₂)_(n)—(CR²R³)_(t),—C(R⁴)═C(R⁵)—, and —C≡C— wherein R² and R³ aremembers independently selected from H, F, substituted or unsubstitutedlower alkyl or substituted or unsubstituted lower heteroalkyl, in whichR² and R³ are optionally joined to form a cyclic structure which is amember selected from the group consisting of cycloalkyl and heterocyclegroups, or R2 and R³ together with the carbon to which they are attachedform —C(O)—; Z represents —O—, —S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—,—C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—, —N(R⁴)C(O)O—, (CR²R³)_(t), and—SO₂N(R⁴)—, wherein R⁴ and R⁵ are members independently selected fromthe group consisting of H, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹ is optionally joined together with either X or R⁴ to form asubstituted or unsubstituted heterocycle; m is an integer from 0 to 2,inclusive; n is an integer from 0 to 3, inclusive; and t is an integerfrom 0 to 2, inclusive.
 16. The method according to claim 15 , whereinsaid voltage-dependent potassium channel is responsible for theM-current.
 17. The method according to claim 15 , wherein saidvoltage-dependent potassium channel comprises KCNQ subunits.
 18. Amethod of treating a central or peripheral nervous system disorder orcondition through modulation of a voltage-dependent potassium channel,said method comprising administering to a subject in need of suchtreatment, an effective amount of a compound having the formula:

wherein Y is a member selected from H, methyl, methoxy,trifluoromethoxy, —CF₃ or halo; V and X are members independentlyselected from H, halo, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, NO₂, CN, CF₃,C(O)NR¹¹R¹² and C(O)R¹³; R¹,R¹¹,R¹² and R¹³ are members independentlyselected from substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, substituted or unsubstitutedcarbocycle, substituted or unsubstituted heterocycle, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹¹ and R¹² optionally can be joined into a ring; Q and W are membersindependently selected from —(CR²R³)_(t)—(CH₂)_(n)—,—(CH₂)_(n)—(CR²R³)_(t), —C(R⁴)═C(R⁵)—, and —C≡C— wherein R² and R³ aremembers independently selected from H, F, substituted or unsubstitutedlower alkyl or substituted or unsubstituted lower heteroalkyl, in whichR² and R³ are optionally joined to form a cyclic structure which is amember selected from the group consisting of cycloalkyl and heterocyclegroups, or R² and R³ together with the carbon to which they are attachedform —C(O)—; Z represents —O—, —S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—,—C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—, —N(R⁴)C(O)O—, (CR²R³)_(t), and—SO₂N(R⁴)—, wherein R⁴ and R⁵ are members independently selected fromthe group consisting of H, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹ is optionally joined together with either X or R⁴ to form asubstituted or unsubstituted heterocycle; m is an integer from 0 to 2,inclusive; n is an integer from 0 to 3, inclusive; and t is an integerfrom 0 to 2, inclusive.
 19. The method according to claim 18 , whereinsaid disorder or condition is selected from the group consisting ofmigraine, ataxia, Parkinson's disease, bipolar disorders, spasticity,mood disorders, brain tumors, psychotic disorders, myokymia, seizures,epilepsy, hearing loss, vision loss, Alzheimer's disease, age-relatedmemory loss, learning deficiencies, motor neuron diseases, and stroke.20. The method according to claim 19 , wherein said disorder orcondition is hearing loss.
 21. The method according to claim 19 ,wherein said disorder or condition is epilepsy or seizures.
 22. A methodof treating a member selected from the group consisting of pain, anxietyand bipolar disorder through modulation of a voltage-dependent potassiumchannel, said method comprising administering to a subject in need ofsuch treatment, an effective amount of a compound having the formula:

wherein Y is a member selected from H, methyl, methoxy,trifluoromethoxy, —CF₃ or halo; V and X are members independentlyselected from H, halo, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, NO₂, CN, CF₃,C(O)NR¹¹R¹² and C(O)R¹³; R¹, R¹¹, R¹² and R¹³ are members independentlyselected from substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, substituted or unsubstitutedcarbocycle, substituted or unsubstituted heterocycle, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹¹ and R¹² optionally can be joined into a ring; Q and W are membersindependently selected from —(CR²R³)_(t)—(CH₂)_(n)—,—(CH₂)_(n)—(CR²R³)_(t), —C(R⁴)═C(R⁵)—, and —C═C— wherein R² and R³ aremembers independently selected from H, F, substituted or unsubstitutedlower alkyl or substituted or unsubstituted lower heteroalkyl, in whichR² and R³ are optionally joined to form a cyclic structure which is amember selected from the group consisting of cycloalkyl and heterocyclegroups, or R² and R³ together with the carbon to which they are attachedform —C(O)—; Z represents —O—, —S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—,—C(O)N(⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—, -N(⁴)C(O)O—, (CR²R³)_(t), and—SO₂N(R⁴)—, wherein R⁴ and R⁵ are members independently selected fromthe group consisting of H, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹ is optionally joined together with either X or R⁴ to form asubstituted or unsubstituted heterocycle; m is an integer from 0 to 2,inclusive; n is an integer from 0 to 3, inclusive; and t is an integerfrom 0 to 2, inclusive.
 23. A composition comprising a pharmaceuticallyacceptable excipient and a compound of the formula:

wherein Y is a member selected from H, methyl, methoxy,trifluoromethoxy, —CF₃ or halo; V and X are members independentlyselected from H, halo, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, NO₂, CN, CF₃,C(O)NR¹¹R¹² and C(O)R¹³; R¹, R¹¹, R¹² and R¹³ are members independentlyselected from substituted or unsubstituted lower alkyl, substituted orunsubstituted lower heteroalkyl, substituted or unsubstitutedcarbocycle, substituted or unsubstituted heterocycle, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹¹ and R¹² optionally can be joined into a ring; Q and W are membersindependently selected from —(CR²R³)_(t)—(CH₂)_(n)—,—(CH₂)_(n)—(CR²R³)_(t), —C(R⁴)═C(R⁵)—, and —C═C— wherein R² and R³ aremembers independently selected from H, F, substituted or unsubstitutedlower alkyl or substituted or unsubstituted lower heteroalkyl, in whichR² and R³ are optionally joined to form a cyclic structure which is amember selected from the group consisting of cycloalkyl and heterocyclegroups, or R² and R³ together with the carbon to which they are attachedform —C(O)—; Z represents —O—, —S(O)_(m)—, —N(R⁴)—, —N(R⁴)C(O)—,—C(O)N(R⁴)—, —C(O)—, —N(R⁴)C(O)N(R⁵)—, —N(R⁴)C(O)O—, (CR²R³)_(t), and—SO₂N(R⁴)—, wherein R⁴ and R⁵ are members independently selected fromthe group consisting of H, substituted or unsubstituted lower alkyl,substituted or unsubstituted lower heteroalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, in whichR¹ is optionally joined together with either X or R⁴ to form asubstituted or unsubstituted heterocycle; m is an integer from 0 to 2,inclusive; n is an integer from 0 to 3, inclusive; and t is an integerfrom 0 to 2, inclusive.